Mitomycin derivatives

ABSTRACT

Mitomycin derivatives of formula &lt;CHEM&gt; wherein W represents a substituted or unsubstituted alkyl having 2 to 19 carbon atoms, a substituted or unsubstituted cycloalkylalkyl, a substituted or unsubstituted alkenyl, a substituted or unsubstituted cycloalkenylalkyl or a substituted or unsubstituted aralkyl; X represents methoxy or amino, or X and W together form -W-X- shown by the formula: &lt;CHEM&gt;  or &lt;CHEM&gt; wherein U und V independently represent a substituted or unsubstituted lower alkyl, lower alkenyl or lower alkoxy group, a substituted or unsubstituted aryl or lower alkenyloxy group, or U and V together form -(CH2)m-CR&lt;3&gt;R&lt;4&gt;-CH2- (wherein m represents an integer of 0 to 1; R&lt;3&gt; and R&lt;4&gt; independently represents hydrogen, a lower alkyl or a substituted or unsubstituted aryl); Y represents hydrogen or methyl; Z represents hydrogen, methyl, a lower alkanoyl or an allyloxycarbonyl; one of R&lt;1&gt; and R&lt;2&gt; represents carbamoyloxymethyl and the other represents hydrogen, or R&lt;1&gt; and R&lt;2&gt; together form methylene (=CH2). The compounds have antitumor activity.

FIELD OF THE INVENTION

The present invention relates to novel mitomycin derivatives having anantitumor activity.

BACKGROUND OF THE INVENTION

As mitomycin derivatives relevant to the present invention where asubstituent is introduced in the methyl group at the 6-position, thereare known mitomycin derivatives where one hydrogen atom of the methylgroup is substituted with deuterium (² H) or tritium (³ H) (JapanesePublished Unexamined Patent Application (hereafter referred as JP-A)70490/89) and mitomycin derivatives where one hydrogen atom of themethyl group is substituted with RO or RS (wherein R representshydrogen, a substituted or unsubstituted alkyl having 1 to 12 carbonatoms, a substituted or unsubstituted alkenyl having 2 to 12 carbonatoms, a substituted or unsubstituted aralkyl or a substituted orunsubstituted aryl) (JP-A 167282/90).

SUMMARY OF THE INVENTION

It is the object of the present invention to provide novel mitomycinderivatives having a substituent in the methyl group at the 6-position.

The present invention relates to mitomycin derivatives represented byformula (I): ##STR1## wherein W represents a substituted orunsubstituted alkyl having 2 to 19 carbon atoms, a substituted orunsubstituted cycloalkylalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted cycloalkenylalkyl or a substituted orunsubstituted aralkyl; X represents methoxy or amino or, X and Wtogether form--W--X-- shown by the formula: ##STR2## wherein U and Vindependently represent a substituted or unsubstituted lower alkyl,lower alkenyl or lower alkoxy group, a substituted or unsubstituted arylor lower alkenyloxy group, or U and V together form--(CH₂)_(m) --CR³ R⁴--CH₂ --(wherein m represents an integer of 0 to 1; R³ and R⁴independently represent hydrogen, a lower alkyl or a substituted orunsubstituted aryl); Y represents hydrogen or methyl; Z representshydrogen, methyl, a lower alkanoyl or an allyloxycarbonyl; one of R¹ andR² represents carbamoyloxymethyl and the other represents hydrogen, orR¹ and R² together form methylene (═CH₂).

DETAILED DESCRIPTION OF THE INVENTION

The compound represented by the formula (I) is referred to herein asCompound (I); and the same shall apply to other compounds having aformula number.

In the definitions of the respective groups in the formula (I), thealkyl is used to mean a straight or branched alkyl group having 2 to 19carbon atoms and the examples of the alkyl include ethyl, propyl,isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, heptyl, octyl,nonyl, decyl, undecyl, dodecyl, octadecyl, nonadecyl, etc.

The cycloalkylalkyl refers to a cycloalkyl group having 4 to 10 carbonatoms and the examples are cyclopropylmethyl, cyclopentylmethyl,cyclohexylmethyl, etc.

The alkenyl refers to a straight or branched alkenyl group having 3 to19 carbon atoms and the examples are allyl, butenyl, isobutenyl,pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl,dodecenyl, octadecenyl, nonadecenyl, etc.

The cycloalkenylalkyl refers to a cycloalkenylalkyl group having 4 to 10carbon atoms and the examples are cyclopentenylmethyl,cyclohexenylmethyl, etc. Examples of the aralkyl include benzyl,phenethyl, benzhydryl, etc. Examples of the aryl include phenyl,naphthyl, etc. The lower alkanoyl refers to a lower alkanoyl grouphaving 1 to 5 carbon atoms and the examples are formyl, acetyl,propionyl, butyryl, isobutyryl, pivaloyl, valeryl, isovaleryl, etc. Thelower alkyl is used to mean a straight or branched alkyl group having 1to 5 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, tert-butyl, pentyl, etc. The lower alkoxy is used to mean astraight or branched alkoxy group having 1 to 5 carbon atoms and theexamples are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy,tert-butoxy, pentyloxy, etc. The alkenyl moiety in the lower alkenyl andlower alkenyloxy refer to a straight or branched alkenyl group having 2to 5 carbon atoms and the examples are propenyl, butenyl, isobutenyl,pentenyl, etc.

In the definitions of the respective groups in the formula (I), one ortwo substituents on the respective groups may be the same or different,and are, for example, lower alkoxy, lower alkanoyl, alkoxycarbonyl,aroyl, oxo, halogen, alkanoylamino, nitro, etc.

The lower alkoxy is as defined for the lower alkoxy described above. Thealkanoyl refers to an alkanoyl group having 1 to 5 carbon atoms and theexamples are formyl, acetyl, propionyl, butyryl, valeryl, etc. Thealkoxycarbonyl refers to an alkoxycarbonyl group having 2 to 5 carbonatoms and the examples are methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, tert-butoxycarbonyl, etc. As the aroyl, there arebenzoyl, p-nitrobenzoyl, naphthoyl, etc. Examples of the halogen includefluorine, chlorine, bromine, etc. The alkanoylamino refers to analkanoylamino group having 2 to 5 carbon atoms and the examples areacetylamino, propionylamino, etc.

Processes for preparing Compound (I) are described below.

Process 1

Compound (I) can be prepared following the steps given below. ##STR3##wherein n represents an integer of 2 or 3; p represents an integer of 1or 2; Y, Z, R¹ and R² are as defined above; W¹ is a group shown by W¹--CH₂ ═W according to the definition of W described above; and Mrepresents a metal atom or a magnesium halide.

The metal atom described above represents sodium, lithium, copper, etc.and examples of the halogen in magnesium halide are chlorine, bromine,iodine, etc.

Step 1:

Compound (IV) can be obtained by reacting Compound (III) with a compoundof the formula W¹ p-M (wherein W¹, M and p are as defined above). Thereaction may also be carried out in the presence of a monovalent coppercatalyst, depending upon the property of the compound W¹ p-M. Examplesof the monovalent copper catalyst are copper (I) iodide, etc. Thesuitable amount of the catalyst is in the range of 0.01 to 1.0equivalent, preferably 0.05 to 0.3 equivalent, based on Compound (III).

Compound (IV) may also be obtained by reacting Compound (III) with acompound of the formula W¹ --H (wherein W¹ is as defined above) in thepresence of a base. As the base, mention may be made of alcoholates,hydroxides, carbonates or hydrogencarbonate of alkali metals or alkalineearth metals, tertiary amines or quaternary ammonium hydroxides, etc.The base may be used in an amount of at least 1 equivalent based onCompound (III).

These processes may be suitably applied depending upon the property ofW¹. As the reaction solvent, there are, for example, ethers such asdiethyl ether, tetrahydrofuran, etc.; halogenated alkane solvents suchas dichloromethane, chloroform, etc.; acetonitrile, dimethylformamide(DMF), dimethylsulfoxide, etc. The solvent may be chosen depending uponreactants and used singly or as admixture. The reaction is carried outgenerally at -78° to 30° C. and completed in 1 minute to 24 hours.

The starting Compound (III) is a known compound described in JP-A70490/89.

Step 2:

Compound (Ia) [in the formula (I), X represents NH₂ ] can be obtained byreacting Compound (IV) with ammonia or ammonium acetate.

Any reaction solvent may be used so long as it is inert to the reactionand can dissolve Compound (IV). As the reaction solvent, there may beused, for example, alcohol type solvents such as methanol, ethanol,etc.; ether type solvents such as diethyl ether, tetrahydrofuran, etc.;halogenated alkane type solvents such as dichloromethane, chloroform,etc.; acetonitrile, DMF, dimethylsulfoxide, etc., singly or incombination. The reaction is carried out generally at 0° to 30° C. andcompleted in an hour to 14 days.

Step 3:

Compound (Ib) [in the formula (I), X represents CH₃ O] can be obtainedby reacting Compound (IV) in the presence of a base or silica gel inmethanol.

As the base, there may be used alcoholates, hydroxides, carbonates orhydrogencarbonates of alkali metals or alkaline earth metal, tertiaryamines or quaternary ammonium hydroxides, etc. The base may be used inan amount of 0.001 to 10 equivalents, preferably 0.01 to 3 equivalents,based on Compound (IV). The reaction is carried out generally at 0° to30° C. and completed in 1 to 24 hours.

As the silica gel, there may be used in an excess amount based onCompound (IV). The reaction is carried out generally 40° C. to theboiling point of the solvent and completed in 1 to 24 hours.

In Steps 2 and 3, deacylation simultaneously occurs in Compound (IV),wherein Z represents a lower alkanoyl, under the reaction conditionsdescribed above to give the corresponding Compound (Ia) and Compound(Ib) wherein Z is a hydrogen atom, respectively.

Process 2

Compound (I) may also be prepared following the steps given below.##STR4## wherein n, W, Y, Z, R¹ and R² are as defined above; Phrepresents phenyl; and Hal represents a halogen atom.

The halogen atom described above refers to chlorine, bromine, iodine,etc.

Step 1:

Compound (VI) can be obtained by reacting Compound (V) with a compoundof the formula W--Hal (wherein Hal and W are as defined above) in thepresence of a base in an inert solvent.

Examples of the reaction solvent are acetone, acetonitrile, dimethylether, tetrahydrofuran, dichloromethane, chloroform, DMF, etc. Thesesolvents may be used singly or as admixture. As the base, mention may bemade of carbonates, hydrogencarbonates or hydroxides of alkali metalssuch as cesium carbonate, potassium carbonate, sodium carbonate, etc.,tertiary amines such as triethylamine, pyridine, etc. or quaternaryammonium hydroxides, etc. The base may be used generally in an excessamount, preferably in 1.5 to 10 equivalents, based on Compound (V). Thereaction may be carried out generally at 0° to 30° C. and completed inan hour to 7 days.

The starting Compound (V) is a known compound described in JP-A14582/91.

Step 2:

Compound (Ia) can be obtained by reacting Compound (VI) with ammonia inthe presence of dimedone in a solvent.

As the solvent, the same solvent as in Step 2 of Process 1 may be used.Dimedone may be used in an excess amount, preferably 2 to 5 equivalentsbased on Compound (VI). The reaction is carried out generally at 0° to30° C. and completed in an hour to 14 days.

Step 3:

Compound (Ib) can be obtained by reacting Compound (VI) in methanol inthe presence of a base and dimedone.

The same solvent and base as used in Step 3 of Process 1 may be used.Dimedone may be used in an excess amount, preferably 2 to 5 equivalentsbased on Compound (V). The amount of the base is used in an excessamount, preferably in the range of 1.01 to 4 equivalents, based ondimedone, and may be varied depending upon a type of the base used. Thereaction is carried out generally at 0° C. to 30° C. and completed in 1to 24 hours.

In Steps 2 and 3, deacylation occurs simultaneously in Compound (VI),wherein Z represents a lower alkanoyl, under the reaction conditionswith dimedone described above to give the corresponding Compound (Ia)and Compound (Ib) wherein Z is a hydrogen atom, respectively.

Process 3 ##STR5## wherein W, X, Y, R¹ and R² are as defined above.

Compound (I) wherein Z is hydrogen, i.e. Compound (Id), can be obtainedby reacting Compound (Ic) wherein Z is an allyloxycarbonyl, with asuitable reducing agent in the presence of a palladium catalyst in asolvent inert to the reaction.

The palladium catalyst is a catalyst which forms a zero valencyhomogeneous active species and there are used, for example, tetrakis(triphenylphosphine) palladium (O) or palladium (II) acetate in thepresence of triphenylphosphine. The amount of the catalyst added is 0.01to 1.0 equivalent, preferably 0.1 to 0.5 equivalent based on Compound(Ic). As the reducing agent, formic acid, triethylammonium formate,tributyltin hydride, triphenyltin hydride, trimethylhydrosilane, sodiumborohydride and the like may be used in an excess amount, based onCompound (Ic). As the solvent, diethyl ether, tetrahydrofuran,acetonitrile, DMF and the like may be used singly or as admixture. Thereaction is carried out at -20° to 80° C., preferably 0° to 30° C. andcompleted in 10 minutes to 10 hours.

Process 4 ##STR6## wherein W, X, Y, R¹ and R² are as defined above.

Compound (I) wherein Z is methyl, i.e. Compound (Ie), can be obtained byreacting Compound (Id), Compound (I) wherein Z is hydrogen, with methyliodide in the presence of a base in a solvent inert to the reaction.

The same solvent and base as used in Step 1 of Process 2 may be used.The reaction is carried out generally at 0° to 120° C. preferably 20° to60° C. and completed in 1 to 30 hours.

Compound (I) wherein X and W are combined together, i.e. Compound (Ih)represented by the formula: ##STR7## wherein R¹, R², Y, Z, U and V areas defined above, can be obtained by the process shown below.

Process 5 ##STR8## wherein n, U, V, Y, Z, R¹ and R² are as definedabove.

Step 1:

Compound (VII) can be obtained by reacting Compound (III) and the saltobtained by treating U--CO--CH₂ --CO--V (wherein U and V are as definedabove) with a base in a solvent inert to the reaction. Depending upon atype of U--CO--CH₂ --CO--V used, Compound (Ih) may further be obtained.

Compound (VII) may also be obtained by reacting Compound (III) withU--CO--CH₂ --CO--V (wherein U and V are as defined above) in thepresence of a base in a solvent inert to the reaction.

Examples of the solvent used in the reaction are ether type solventssuch as diethyl ether, tetrahydrofuran, etc.; halogenated alkane typesolvents such as dichloromethane, chloroform, etc.; acetonitrile, DMF,dimethylsulfoxide, etc. The suitable solvent may be chosen dependingupon the properties of reactants and used singly or as admixture. As thebase, there may be used hydrides, alcoholate, carbonates orhydrogencarbonates of alkali metals or alkaline earth metals; tertiaryamines or quaternary ammonium hydroxides. The base may be used in anamount of 1 equivalent or more. The reaction is carried out generally at-78° to 30° C. and completed in a minute to 24 hours.

Step 2:

Compound (Ih) can be obtained by dissolving Compound (VII) in a solvent,adsorbing onto silica gel and allowing to stand.

As the solvent, mention may be made of halogenated alkane solvents suchas dichloromethane, chloroform, etc. The reaction is carried outgenerally at 0° to 40° C. and completed in an hour to 7 days.

Process 6

Compound (Ih) wherein Z is hydrogen i.e. Compound (Iha), can also beobtained by reacting Compound (Ihc) where Z is a lower alkanoyl, in thepresence of a base in methanol.

Examples of the base which can be used in the reaction are ammonia,alcoholates, hydroxides, carbonates or hydrogencarbonates of alkalimetals or alkaline earth metals; tertiary amines or quaternary ammoniumhydroxides. The base is used in a catalytic amount or in an excessamount based on Compound (Ihc), depending upon a type of the base. Thereaction is carried out at 0° to 30° C. and completed in 1 to 24 hours.

Process 7

Compound (Ih) wherein Z is hydrogen, i.e. Compound (Iha), can beprepared by reacting Compound (Ihd) wherein Z is an allyloxycarbonyl,with a suitable reducing agent in the presence of a palladium catalystin a solvent inert to the reaction. ##STR9## wherein U, V, Y, R¹ and R²are as defined above.

The palladium catalyst used is a catalyst that forms a zero valencyhomogeneous active species and there are used, for example, tetrakis(triphenylphosphine) palladium (O) or palladium (II) acetate in thepresence of triphenylphosphine. The amount of the catalyst added is 0.01to 1.0 equivalent, preferably 0.1 to 0.5 equivalent based on Compound(Ihd).

As the reducing agent, formic acid, triethylammonium formate,tributyltin hydride, triphenyltin hydride, trimethylhydrosilane and thelike may be used in an excess amount, based on Compound (Ihd).

As the solvent, diethyl ether, tetrahydrofuran, acetonitrile, DMF andthe like may be used singly or as admixture.

The reaction is carried out at -20° to 80° C., preferably 0° to 30° C.and completed in 10 minutes to 10 hours.

The intermediates and the desired products in the process describedabove may be isolated and purified by purification techniquesconventionally used in organic synthetic chemistry, for example,neutralization, filtration, extraction, washing, drying, concentration,recrystallization, various chromatographies, etc. In the case of theintermediates, they can be applied directly to the subsequent stepwithout any further purification. Furthermore, Compounds (I) may beobtained in the form of addition products with water or varioussolvents, which are also within the scope of the present invention.

Specific examples of Compounds (I) obtained by the processes describedabove are shown in Tables 1 and 2.

                                      TABLE 1    __________________________________________________________________________     ##STR10##    Compound          (Example No.)                  W           X   Y  Z  R.sup.1 R.sup.2    __________________________________________________________________________    1      (1)    CH.sub.2 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    2      (2)    CH.sub.2 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     CH.sub.3                                        CH.sub.2 OCONH.sub.2                                                H    3      (3)    CH.sub.2 CH.sub.3                              NH.sub.2                                  H  CH.sub.3                                        H       CH.sub.2 OCONH.sub.2    4      (4)    (CH.sub.2).sub.2 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    5      (5)    CH.sub.2 CHCH.sub.2                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    6      (6)    (CH.sub.2).sub.4 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    7      (7)    CH.sub.2 Ph NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    8      (8)    (CH.sub.2).sub.2 NO.sub.2                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    9      (9)                   ##STR11##  NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    10    (10)                   ##STR12##  OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    11    (11)                   ##STR13##      CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    15    (15)    CH.sub.2 CH.sub.3                              OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    16    (16)    CH.sub.2 CH.sub.3                              OCH.sub.3                                  H  CH.sub.3                                        H       CH.sub.2 OCONH.sub.2    17    (17)    CH.sub.2 CH(CH.sub.3).sub.2                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    18    (18)    CH.sub.2 CH(CH.sub.3).sub.2                              OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    19    (19)    CH.sub.2 CH(CH.sub.3).sub.2                              OCH.sub.3                                  H  CH.sub.3                                        H       CH.sub.2 OCONH.sub.2    20    (20)    (CH.sub.2).sub.4 CH.sub.3                              OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    21    (21)    (CH.sub.2).sub.4 CH.sub.3                              OCH.sub.3                                  H  CH.sub.3                                        H       CH.sub.2 OCONH.sub.2    22    (22)    CH.sub.2 Ph OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    23    (23)    CH.sub.2 Ph OCH.sub.3                                  H  CH.sub.3                                        H       CH.sub.2 OCONH.sub.2    24    (24)    (CH.sub.2).sub.10 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    25    (25)    (CH.sub.2).sub.10 CH.sub.3                              OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    26    (26)    (CH.sub.2).sub.18 CH.sub.3                              NH.sub.2                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    27    (27)    (CH.sub.2).sub.18 CH.sub.3                              OCH.sub.3                                  CH.sub.3                                     H  CH.sub.2 OCONH.sub.2                                                H    28    (28)                   ##STR14##  OCH.sub.3                                  CH.sub.3                                     CH.sub.3                                        CH.sub.2 OCONH.sub.2                                                H    __________________________________________________________________________

                                      TABLE 2    __________________________________________________________________________     ##STR15##    Compound          (Example No.)                  U     V      Y  Z  R.sup.1 R.sup.2    __________________________________________________________________________    12    (12)    CH.sub.3                        OCH.sub.2 CH.sub.3                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    13    (13)    CH.sub.2 C(CH.sub.3).sub.2 CH.sub.2                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    14    (14)    (CH.sub.2).sub.3                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    29    (29)    CH.sub.3                        CH.sub.3                               CH.sub.3                                  CH.sub.3                                     CH.sub.2 OCONH.sub.2                                             H    30    (30)    CH.sub.3                        Ph     CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    31    (31)    CH.sub.3                        Ph     CH.sub.3                                  CH.sub.3                                     CH.sub.2 OCONH.sub.2                                             H    32    (32)    CH.sub.3                        OCH.sub.2 CH.sub.3                               CH.sub.3                                  CH.sub.3                                     CH.sub.2 OCONH.sub.2                                             H    33    (28)    ρ-NO.sub.2 Ph                        OCH.sub.2 CH.sub.3                               CH.sub.3                                  CH.sub.3                                     CH.sub.2 OCONH.sub.2                                             H    34    (33)    CH.sub.2 C(CH.sub.3).sub.2 CH.sub.2                               H  CH.sub.3                                     H       CH.sub.2 OCONH.sub.2    35    (34)    (CH.sub.2).sub.3                               H  CH.sub.3                                     H       CH.sub.2 OCONH.sub.2    36    (35)    CH.sub.2 CH(CH.sub.3)CH.sub.2                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    37    (35)    CH.sub.2 CH(CH.sub.3)CH.sub.2 *.sup.1                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    38    (36)    CH.sub.2 CH(ρ-CH.sub.3 OPh)CH.sub.2                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    39    (36)    CH.sub.2 CH(ρ-CH.sub.3 OPh)CH.sub.2 *.sup.2                               CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    40    (37)                   ##STR16##   CH.sub.3                                  H  CH.sub.2 OCONH.sub.2                                             H    __________________________________________________________________________     *.sup.1 Diastereomer of Compound 36     *.sup.2 Diastereomer of Compound 38

In the following, the antitumor activity of representative Compound (I)is specifically shown with reference to the experiments.

Experiment 1 Growth inhibition test on HeLaS₃ cells

HeLaS₃ cells were suspended in an MEM medium containing 10% fetal calfserum and 2 mM glutamine at a density of 3×10⁴ cells/ml, and 0.1 ml ofthe cell suspension was put into each well of a 96-well microtiterplate.

After culturing at 37° C. overnight in a carbon dioxide gas incubator,0.05 ml each of a test compound appropriately diluted with the mediumwas added to each well. The cells were further cultured for an hour inthe carbon dioxide gas incubator, and the culture supernatant wasremoved. The residue was washed once with a phosphate buffered saline[PBS (-)] and 0.1 ml each of a fresh medium was added to each well. Thecells were further incubated at 37° C. for 72 hours in the carbondioxide gas incubator. After removal of the culture supernatant, 0.1 mleach of the culture medium containing 0.02% Neutral Red was added toeach well, and the cells were further incubated at 37° C. for an hour inthe carbon dioxide gas incubator to stain the cells.

After removal of the culture supernatant, each well was washed once witha physiological saline, and the dye was extracted with 0.001N HCl/30%ethanol. Absorbance of the extract at 550 nm was measured with amicroplate reader. The cell growth inhibition percentage was calculatedaccording to the following equation from the absorbance of the extractof the cells treated with the test compound in various concentrationsand that of the untreated cells. ##EQU1##

From the cell growth inhibition percentage thus obtained, theconcentration of the test compound which inhibits cell growth by 50%,i.e. IC₅₀ value, was determined.

The results are shown in Table 3.

                  TABLE 3    ______________________________________    Compound      IC.sub.50 (mM)    ______________________________________    1             3.0    7             2.3    9             1.5    10            1.6    13            0.084    14            0.065    15            0.017    16            0.30    18            0.20    20            0.087    22            0.063    23            0.52    29            0.36    31            0.31    32            0.10    36            0.093    37            0.076    38            0.14    39            0.43    40            0.53    ______________________________________

Experiment 2 Anti-tumor activity to sarcoma 180 solid tumor

5×10⁶ Sarcoma 180 cells were abdominally implanted into a ddY mouse andthe cells were collected from the ascites of the mouse 7 days after theimplantation. The cells were washed once with a sterilized physiologicalsaline solution and then suspended in a sterilized physiological salinesolution to prepare a cell suspension of 5×10⁷ cells/ml.

0.1 ml of the cell suspension was subcutaneously implanted into theright axillary space of male ddY mice weighing 20±2 g.

A test compound was dissolved in a physiological saline solution or apolyoxyethylene sorbitan monolaurate-containing physiological salinesolution, and 0.1 to 0.2 ml of the solution was intravenously injectedto 5 mice as one group 24 hours after the implantation of the tumorcells.

The anti-tumor activity of the test compound was determined by measuringthe major axis (a) and the minor axis (b) of the tumor 7 days after theimplantation of tumor cells, and the value of (a×b² /2), whichcorresponds to the volume of the tumor, was calculated. The intendedanti-tumor activity is represented by a ratio of T/C, in which Cindicates the tumor volume of mice of the control group to which no testcompound was administered and T indicates the tumor volume of mice ofthe test group to which the test compound was administered. T/C at eachdose given was plotted on the vertical axis and a dose is shown by alogarithmic scale on the horizontal axis. The relationship between thedose and T/C was determined to be a straight line by the least squaresmethod. From the regression formula of the straight line thus obtained,the dose showing T/C=0.5 was calculated to give ED₅₀.

The results are shown in Table 4.

                  TABLE 4    ______________________________________    Compound      ED.sub.50 (mg/kg)    ______________________________________    1             6.7    7             18    9             8.0    12            0.70    13            1.3    14            0.84    15            2.8    16            14    30            5.8    36            1.3    37            0.93    38            12    ______________________________________

Experiment 3 Acute toxicity

A test compound was intravenously injected once to five ddY mice as onegroup. After the administration, the mice were observed for 14 days anddeaths were recorded. LD₅₀ was calculated from the death rate of eachgroup to which the test compound was administered according to theBehrens-Kaerber's method.

The results are shown in Table 5.

                  TABLE 5    ______________________________________    Compound      LD.sub.50 (mg/kg)    ______________________________________    1             38    7             >50    9             30    12            5.2    13            2.9    14            2.2    15            8.2    16            25    36            4.4    ______________________________________

The compounds obtained by the present invention are useful as anti-tumoragents, which can be used directly as such, or in various dosage forms.For example, where Compound (I) is used in the form of an injection, itmay be dissolved in a diluent conventionally used in the art, such as aphysiological saline, or glucose, lactose or mannitol solution forinjection. Alternatively, Compound (I) may be freeze-dried according toa conventional manner to give a product for injection or may be preparedinto injectable powder by adding sodium chloride thereto. In addition,the injection may also contain an auxiliary agent such as polyethyleneglycol, HCO-60 (surfactant manufactured by Nikko Chemical Co., Ltd.), aswell as a carrier such as ethanol and/or liposome or cyclodextrin. Theseinjections are generally used for intravenous administration but mayalso be used for intra-arterial, intraperitoneal or intra-thoracialadministration.

Compound (I) may also be administered orally by mixing with anappropriate excipient, a disintegrator, a binder, a lubricant, etc. in aconventional manner to prepare a tablet, a granule, a powder or a syrup.Furthermore, Compound (I) may be mixed with a conventionally usedcarrier and formed into a suppository for rectum administration.

Dosage may appropriately vary depending upon the administration route,the kind of Compound (I), and the age and condition of a patient. Theadministration route, may also be varied according to the condition of apatient and the dosage. For example, Compound (I) can be intermittentlyadministered in a dose of 0.06 to 6 mg/kg, once a week or once every 3weeks.

Hereafter the present invention is described with reference to thefollowing examples.

Physicochemical data of each compound were obtained using the followingdevices.

    ______________________________________    TLC:           silica gel Art 5715                   (manufactured by Merck Inc.)    MS: Hitachi    M-80B (determined by El or Sl                   method)    JEOL, Ltd.     JMS-D300 (determined by FAB                   method)    IR: Nippon Bunko                   IR-810 (measured by the KBr                   method)    .sup.1 H-NMR:    Bruker         AM400 (400 MHz)    JEOL, Ltd.     JNM-GX270 (270 MHz)    VARIAN         EM390 (90 MHz)    ______________________________________

EXAMPLES EXAMPLE 1 Synthesis of 6-demethyl-6-ethylmitomycin C (Compound1)

After 55.9 mg ofIa-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycinA (JP-A 70940/89) was dissolved in 1.0 ml of anhydrous acetonitrile, 100μl of ethyl iodide and 65 mg of cesium carbonate were added to thesolution. The mixture was stirred at 20° C. for 45 hours. After thereaction mixture was diluted with dichloromethane, and washedsuccessively with a phosphate buffer (pH 4), saturated aqueous sodiumhydrogencarbonate solution and saturated aqueous sodium chloridesolution. The reaction mixture was then dried over anhydrous sodiumsulfate. After the desiccant was removed by filtration, the solvent wasdistilled off to give the crude product ofla-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-ethyl-6-phenylselenomitomycinA.

The whole amount of the resulting crude product was dissolved in 2.0 mlof a 6.1M solution of ammonia in methanol and 50 mg of dimedone wasadded to the solution. The mixture was stirred at 20° C. for 3 hours anda half. The reaction mixture was diluted with dichloromethane and thenwashed successively with a phosphate buffer (pH 4), saturated aqueoussodium hydrogencarbonate solution and saturated aqueous sodium chloridesolution. After the reaction mixture was dried over anhydrous sodiumsulfate, the solvent was distilled off. The residue obtained waspurified by preparative TLC (silica gel; chloroform:methanol=9:1) togive a violet fraction containing the desired product. This fraction wasextracted with the developing solvent. After the solvent was distilledoff, the residue was dissolved in a small amount of chloroform. n-Hexanewas added to the solution to give a powder. The solvent was distilledoff and the residue was thoroughly dried in vacuo to give 6.0 mg (yield17%) of Compound 1 as a violet powder.

TLC: Rf=0.35 (chloroform:methanol=9:1)

EI-MS: m/e 348 (M⁺); C₁₆ H₂₀ N₄ O₅ =348

IR (cm⁻¹): 3430, 3330, 2930, 1710, 1650, 1600, 1540, 1450, 1340, 1260,1220, 1070

¹ H-NMR (400 MHz, pyridine -d₅) δ(ppm): 1.08 (t, J=7.4 Hz, 3H), 2.10(br, 1H), 2.61(q, J=7.4 Hz, 2H), 2.74 (dd, J=1.7 & 4.4 Hz, 1H), 3.13 (d,J=4.2 Hz, 1H), 3.21 (s, 3H), 3.61 (dd, J=1.7 & 12.6 Hz, 1H), 4.03 (dd,J=4.2 & 11.1 Hz, 1H), 4.58 (d, J=12.6 Hz, 1H), 5.08 (bt, J=11.1 Hz, 1H),5.43 (dd, J=4.2 & 10.3 Hz, 1H), 7.60 (br, 2H), 7.67 (br, 2H)

EXAMPLE 2 Synthesis of 6-demethyl-6-ethylporfiromycin (Compound 2)

Compound 1 (50.0 mg) obtained in EXAMPLE 1 was dissolved in acetone and,19.4 mg of potassium carbonate and 0.5 ml of methyl iodide were added tothe solution. The mixture was stirred at 20° C. for 23 hours. Thesolvent was distilled off and the residue was purified by columnchromatography (silica gel; chloroform:methanol=30:1) to give a violetfraction. From this fraction the solvent was distilled off and a smallamount of chloroform was added thereto. n-Hexane was added to themixture to give a powder. The powder was then thoroughly dried in vacuoto give 30.0 mg (yield 58%) of Compound 2 as a violet powder.

TLC: Rf=0.37 (chloroform:methanol=9.1)

FAB-MS (m/z): 363 (M⁺ +1); C₁₇ H₂₂ N₄ O₅ =362

IR (cm⁻¹): 3430, 3320, 3200, 2950, 2870, 1720, 1600, 1570, 1550, 1450,1340, 1210, 1110, 1070

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 1.09 (t, J=7.4 Hz, 3H), 2.15 (dd,J=2.1 & 4.8 Hz, 1H), 2.24 (s, 3H), 2.54 (d, J=4.6 Hz, 1H), 2.63 (q,J=7.6 Hz, 2H), 3.18(s, 3H), 3.54 (dd, J=2.1 & 12.8 Hz, 1H), 4.01 (dd,J=4.2 & 11.5 Hz, 1H), 4.52 (d, J=12.9 Hz, 1H), 4.83 (dd, J=10.5 & 11.2Hz, 1H), 5.39 (dd, J=4.3 & 10.4 Hz, 1H), 7.4˜8.0 (br, 4H)

EXAMPLE 3 Synthesis of 6-demethyl-6-ethylmitomycin D (Compound 3)

After 1.19 g (purity 60%) of Compound c obtained in REFERENCE EXAMPLE 3was dissolved in 20 ml of anhydrous tetrahydrofuran, 18 ml of a 0.25Msolution of lithium dimethyl cuprate in tetrahydrofuran was addeddropwise at -78° C. over 30 minutes. The mixture was stirred at -78° C.for 30 minutes. Saturated aqueous ammonium chloride solution was addedto the reaction mixture. The mixture was stirred and then extracted withchloroform. The organic layer was washed with saturated aqueous sodiumchloride solution, and dried over anhydrous sodium sulfate, and then thesolvent was distilled off. The residue obtained was dissolved in a smallamount of chloroform and n-hexane was added to the solution to give apowder. The solvent was again distilled off to give the crude product of7-demethoxy-6-demethyl-6-ethyl-7-ethylenedioxy-6,7-dihydromitomycin B asa brown powder.

The whole amount of the crude product obtained was dissolved in 50 ml ofmethanol and 10 ml of a 6.8M solution of ammonia in methanol. Thesolution was allowed to stand at 20° C. for 9 hours and a half. Afterthe solvent was distilled off, the residue obtained was purified bycolumn chromatography (silica gel; chloroform:methanol=25:1) to give aviolet fraction. This fraction was treated in a conventional manner togive 76.4 mg (yield 12%) of Compound 3 as a violet powder.

TLC: Rf=0.28 (chloroform:methanol=9:1)

EI-MS: m/z 348 (M⁺); C₁₆ H₂₀ N₄ O₅ =348

IR (cm⁻¹): 3420, 3330, 3270, 3200, 2950, 2870, 1710, 1700, 1600, 1560,1550, 1540, 1450, 1420, 1350, 1060

¹ H-NMR (90 MHz, pyridine-d₅) δ(ppm): 1.04 (t, J=7.5 Hz, 3H), 2.13(s,3H), 2.23 (dd, J=1.6 & 4.8 Hz, 1H), 2.48 (d, J=4.8 Hz, 1H), 2.57(q,J=7.5 Hz, 2H), 3.69 (dd, J=1.8 & 13 Hz, 1H), 4.27 (dd, J=3.7 & 10 Hz,1H), 4.48 (d, J=13 Hz, 1H), 5.25(t, J=10 Hz, 1H), 5.55 (dd, J=3.7 & 10Hz, 1H), 7.3˜7.6 (br, 4H), 8.23 (s, 1H)

EXAMPLE 4 Synthesis of 6-demethyl-6-n-propylmitomycin C (Compound 4)

To a solution of 430 mg of Compound a obtained in REFERENCE EXAMPLE 1 in20 ml of anhydrous tetrahydrofuran was added 50 mg of copper (I) iodidefollowed by the dropwise addition of 1.0 ml of a 3.0M solution ofethylmagnesium bromide in diethyl ether at -78° C. over 20 minutes. Themixture was stirred at -78° C. for further 40 minutes. The yellow orangereaction mixture was diluted with a phosphate buffer (pH 4). After theresultant reddish pink organic layer was separated, the aqueous layerwas extracted with dichloromethane. The organic layer was washedsuccessively with saturated aqueous sodium hydrogencarbonate solutionand saturated aqueous sodium chloride solution and then dried overanhydrous sodium sulfate. The solvent was then distilled off. Theresidue was purified by column chromatography (silica gel;chloroform:methanol=20:1) to give a yellowish pink fraction containingthe desired product. This fraction was treated in a conventional mannerto give 112.8 mg (yield 24%) oflacetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-propylmitomycinA as a yellowish pink powder.

The whole amount of the product obtained above was dissolved in 10 ml ofmethanol and 1.0 ml of a 6.1M solution of ammonia in methanol. Theresulting solution was allowed to stand at 20° C. for 22 hours and 20minutes and at 5° C. for 40 hours. The reaction mixture was concentratedin vacuo. The residue obtained was purified by preparative TLC (silicagel; chloroform:methanol=9:1) to give a violet fraction containing thedesired product. This fraction was extracted, crystallized withdichloromethane and fully dried in vacuo in a conventional manner togive 12.7 mg (yield, 14%) of Compound 4 as violet needles. The filtratewas treated by conventional procedure to give a powder and dried invacuo to give 13.9 mg (yield, 16%) of the product as a violet powder.

TLC: Rf=0.43 (chloroform:methanol=9:1)

EI-MS: m/z 362 (M⁺); C₁₇ H₂₂ N₄ O₅ =362

IR (cm⁻¹): 3430, 3320, 3280, 3200, 2950, 2860, 1710, 1600, 1560, 1550,1450, 1370, 1340, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 0.84 (t, J=7.3 Hz, 3H),1.45˜1.65 (m, 2H), 1.9˜2.2 (br, 1H), 2.56 (bt, J=7.7 Hz, 2H), 2.73 (bs,1H), 3.13 (bs, 1H), 3.21 (s, 3H), 3.60 (bd, J=13.4 Hz, 1H), 4.03 (dd,J=4.1 & 11.1 Hz, 1H), 4.57 (d, J=12.7 Hz, 1H), 5.0˜5.2 (br, 1H), 5.42(dd, J=4.3 & 10.4 Hz, 1H), 7.3˜7.8 (br, 4H)

EXAMPLE 5 Synthesis of 6-allyl-6-demethyl-mitomycin C (Compound 5)

After Compound b (56.0 mg) obtained in REFERENCE EXAMPLE 2 was dissolvedin 1.0 ml of anhydrous acetone, 100 μl of allyl iodide and 65 mg ofcesium carbonate were added to the solution. The mixture was stirred at20° C. for 40 hours and a half. The reaction mixture was diluted withchloroform, washed successively with a phosphate buffer (pH 4),saturated aqueous sodium hydrogencarbonate solution and saturatedaqueous sodium chloride solution, and then dried over anhydrous sodiumsulfate. After the solvent was distilled off, the residue was purifiedby preparative TLC (silica gel; chloroform:methanol=9:1) to give ayellow fraction. This fraction was treated in a conventional manner togive 30.1 mg (yield 50%) ofla-acetyl-6-allyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycinA as a yellow powder. The purity of the compound was about 70% by ¹H-NMR.

23.9 mg of the product obtained above was dissolved in 2.0 ml of a 6.1Msolution of ammonia in methanol and 40 mg of dimedone was added to thesolution. The mixture was stirred at 20° C. for 20 hours. After dilutingthe reaction mixture with dichloromethane, the organic layer was washedwith saturated aqueous sodium chloride solution. The reaction mixturewas then dried over anhydrous sodium sulfate. After the solvent wasdistilled off, the residue was purified by preparative TLC (silica gel;chloroform:methanol=9:1) to give a violet fraction containing thedesired product. This fraction was treated in a conventional manner togive 4.6 mg (yield, about 46%) of Compound 5 as a violet powder.

TLC: Rf=0.25 (chloroform:methanol=9:1)

FAB-MS (m/z): 361 (M⁺ +1); C₁₇ H₂₀ N₄ O₅ =360

IR (cm⁻¹): 3420, 3320, 3200, 2920, 2850, 1720, 1710, 1600, 1560, 1550,1450, 1440, 1370, 1340, 1230, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 2.07 (bs, 1H), 2.73 (bs, 1H),3.16 (bs, 1H), 3.21 (s, 3H), 3.3˜3.5 (m, 2H), 3.59 (bd, J=12.3 Hz, 1H),4.04 (dd, J=4.1 & 11.1 Hz, 1H), 4.56 (d, J=12.6 Hz, 1H), 4.99 (dd, J=1.5& 9.9 Hz, 1H), 5.12 (bt, J=11 Hz, 1H), 5.21 (dd, J=1.7 & 19.0 Hz, 1H),5.43 (dd, J=4.2 & 10.4 Hz, 1H), 5.85˜6.02 (m, 1H), 7.3˜7.8 (br, 4H)

EXAMPLE 6 Synthesis of 6-demethyl-6-n-pentylmitomycin C (Compound 6)

After 190 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 6.0 ml of anhydrous tetrahydrofuran, 5.3 ml of a 0.094M solution oflithium di-n-butyl cuprate in tetrahydrofuran was added dropwise at -78°C. over 30 minutes. The mixture was stirred at -78° C. for 50 minutes.Saturated aqueous ammonium chloride solution was added to the reactionmixture. The mixture was stirred and then extracted with chloroform. Theorganic layer was washed with saturated aqueous sodium chloride solutionand dried over anhydrous sodium sulfate and then the solvent wasdistilled off. The residue obtained was purified by columnchromatography (silica gel; chloroform:methanol=50:1 to 30:1) to give ared fraction. This fraction was treated in a conventional manner to give74.4 mg (yield, 34%) of aIa-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-pentylmitomycinA as a pink powder.

The whole amount of the product obtained above was dissolved in 4.8 mlof a 6.8M solution of ammonia in methanol. The resulting solution wasallowed to stand at 20° C. for 23 hours. After the solvent was distilledoff, the residue obtained was purified by column chromatography (silicagel; chloroform:methanol=30:1) to give a violet fraction. This fractionwas treated in a conventional manner to give 28.2 mg (yield 46%) ofCompound 6 as a violet powder.

TLC: Rf=0.37 (chloroform:methanol=9:1)

SI-MS (m/z): 392 (M⁺ +2); C₁₉ H₂₆ N₄ O₅ =390

IR (cm⁻¹): 3400, 3320, 2930, 1700, 1650, 1590, 1540, 1450, 1340, 1270,1220, 1070

¹ H-NMR (400 MHz, chloroform -d₁) δ(ppm): 0.65 (br, 1H), 0.89 (t, J=6.9Hz, 3H), 1.26˜1.40 (m, 6H), 2.20˜2.24 (m, 2H), 2.82 (br, 1H), 2.89 (d,J=4.4 Hz, 1H), 3.22 (s, 3H), 3.51 (bd, J=12.8 Hz, 1H), 3.62 (dd, J=4.4 &10.6 Hz, 1H), 4.25 (d, J=12.8 Hz, 1H), 4.54 (bt, J=10.6 Hz, 1H), 4.69(br, 2H), 4.71 (dd, J=4.4 & 10.6 Hz, 1H), 5.24 (br, 2H)

EXAMPLE 7 Synthesis of 6-benzyl-6-demethylmitomycin C (Compound 7)

After Compound b (28.1 mg) obtained in REFERENCE EXAMPLE 2 was dissolvedin 0.50 ml of anhydrous acetone, 60 μl of benzyl bromide and 35 mg ofcesium carbonate were added to the solution. The mixture was stirred at20° C. for 13 hours. After the reaction mixture was diluted withchloroform, the dilution was washed successively with saturated aqueoussodium hydrogencarbonate solution and saturated aqueous sodium chloridesolution. The reaction mixture was then dried over anhydrous sodiumsulfate. After the solvent was distilled off, the residue was purifiedby preparative TLC (silica gel; chloroform:acetonitrile:n-hexane=5:5:1)to give a yellow fraction. This fraction was treated in a conventionalmanner to give 15.3 mg of the crude product containingIa-acetyl-6-benzyl-7-demethoxy-6-demethyl-6,7-dihydro-6-phenylselenomitomycinA as a yellow powder.

The whole amount of the crude product obtained was dissolved in 1.0 mlof a 6.1M solution of ammonia in methanol and 10 mg of dimedone wasadded. The reaction mixture was stirred at 20° C. for 40 minutes andthen diluted with dichloromethane. Subsequently, the dilution was washedwith saturated aqueous sodium chloride solution and dried over anhydroussodium sulfate. After the solvent was distilled off, the residueobtained was purified by preparative TLC (silica gel;chloroform:methanol=9:1) to give a violet fraction containing thedesired product. This fraction was treated in a conventional manner togive 4.5 mg (yield 22%) of Compound 7 as a violet powder.

TLC: Rf=0.44 (chloroform:methanol=9:1)

FAB-MS (m/z): 411 (M⁺ +1); C₂₁ H₂₂ N₄ O₅ =410

IR (cm⁻¹): 3420, 3320, 2920, 1720, 1710, 1600, 1560, 1550, 1450, 1440,1370, 1340, 1220, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 2.05 (bs, 1H), 2.72 (bs, 1H),3.13 (bs, 1H), 3.19 (s, 3H), 3.58 (bd, J=12.3 Hz, 1H), 3.96 (d, J=15.4Hz, 1H), 4.04 (d, J=15.4 Hz, 1H), 4.04 (dd, J=4.4 & 11.4 Hz, 1H), 4.55(d, J=12.6 Hz, 1H), 5.12 (bt, J=11 Hz, 1H), 5.42 (dd, J=4.3 & 10.3 Hz,1H), 7.08˜7.26 (m, 3H), 7.41˜7.57 (m, 2H), 7.4˜7.8 (br, 2H), 7.7 (bs,2H)

EXAMPLE 8 Synthesis of 6-demethyl-6-(2-nitroethyl)mitomycin C (Compound8)

Compound a (1.00 g) obtained in REFERENCE EXAMPLE 1 was dissolved in 50ml of dichloromethane, and 660 mg of potassium carbonate and 2.5 ml ofnitromethane were added to the solution. The mixture was stirred at 20°C. for 2 hours and a half. The reaction mixture was washed withsaturated aqueous sodium chloride solution and then dried over anhydroussodium sulfate. The solvent was then distilled off. The residue obtainedwas purified by column chromatography (silica gel;chloroform:methanol=20:1) to give a yellow fraction. This fraction wastreated in a conventional manner to give 547.7 mg (yield 48%) ofIa-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-(2-nitroethyl)mitomycinA as a yellow powder.

The whole amount of the product above was dissolved in 50 ml ofanhydrous tetrahydrofuran and the solution was allowed to stand at 20°C. for 70 hours under anhydrous ammonia atmosphere. The solvent wasdistilled off from the reaction mixture. The residue obtained waspurified by column chromatography (silica gel; chloroform:methanol=20:1to 10:1) to give a violet fraction. From this fraction the solvent wasdistilled off and the residue was crystallized frommethanoldichloromethane to give 123.6 mg (yield 28%) of Compound 8 asviolet prisms.

TLC: Rf=0.22 (chloroform:methanol=9:1)

SI-MS (m/z): 395 (M⁺ +2); C₁₆ H₁₉ N₅ O₇ =393

IR (cm⁻¹): 3420, 3340, 2940, 1710, 1660, 1600, 1560, 1540, 1480, 1450,1370, 1330, 1190, 1170, 1060

¹ H-NMR (400 MHz, pyridine -d₅) δ(ppm): 2.23 (br, 1H), 2.77 (br, 1H),3.15 (br, 1H), 3.22 (s, 3H), 3.27˜3.40 (m, 2H), 3.60 (bd, J=12.3 Hz,1H), 4.03 (dd, J=4.2 & 11.1 Hz, 1H), 4.54 (d, J=12.8 Hz, 1H), 4.70 (t,J=7.6 Hz, 2H), 5.09 (bt, J=10.3 Hz, 1H), 5.42 (dd, J=4.2 & 10.3 Hz, 1H),7.63 (br, 2H), 8.20 (br, 2H)

EXAMPLE 9 Synthesis of Compound 9

422 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolved in 30ml of anhydrous tetrahydrofuran and 152 mg of dimedone and 100 μl ofanhydrous triethylamine were added to the solution followed by stirringat ambient temperature for 45 minutes. The reaction mixture was washedwith saturated aqueous sodium chloride solution and then dried overanhydrous sodium sulfate. The solvent was then distilled off. Theresidue was purified by column chromatography (silica gel;chloroform:methanol=20:1 to 10:1) to give violet to yellow fractions.The fractions were treated in a conventional manner to give the crudeproduct (378.7 mg) containingIa-acetyl-7-demethoxy-6-demethyl-6-[(4,4-dimethylcyclohexane-2,6-dion-1-yl)-7-ethylenedioxy-6,7-dihydromethyl]mitomycin A as a violet powder.

The product (241.2 mg) obtained above was dissolved in 10 ml of a 6.1Msolution of ammonia in methanol and 100 mg of dimedone was added to thesolution. The mixture was stirred at 20° C. for 3 hours. The reactionmixture was concentrated in vacuo and chloroform was added to theconcentrate to form a solution. The solution was washed with saturatedaqueous sodium chloride solution. The aqueous layer was allowed to standfor a while and extracted with chloroform after the solution was turnedviolet. The organic layers were combined and dried over anhydrous sodiumsulfate. The solvent was distilled off. The residue obtained waspurified by preparative TLC (silica gel; chloroform:methanol=9:1) togive a green fraction. This fraction was treated in a conventionalmanner to give 51.6 mg (yield, 17%) of the product as a green powder.

TLC:Rf=0.47 (chloroform:methanol=9:1)

FAB-MS (m/z): 474 (M⁺ +2); C₂₃ H₂₈ N₄ O₇ =472

IR (cm⁻¹): 3400, 3170, 2950, 2880, 1720, 1710, 1610, 1560, 1520, 1450,1350, 1240, 1220, 1160, 1060

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 0.92 (s, 6H), 2.11 (bs, 1H),2.33 (bs, 4H), 2.75 (bs, 1H), 3.12 (bs, 1H), 3.18 (s, 3H), 3.37 (d,J=15.0 Hz, 1H), 3.45 (d, J=15.0 Hz, 1H), 3.58 (bd, J=12.7 Hz, 1H), 3.96(dd, J=4.3 & 11.1 Hz, 1H), 4.45 (d, J=12.9 Hz, 1H), 5.02 (bt, J=11 Hz,1H), 5.33 (dd, J=4.3 & 10.4 Hz, 1H), 7.4˜7.8 (br, 2H), 8.62 (bs, 1H),8.95 (bs, 1H), 14.5 (br, 1H)

EXAMPLE 10 Synthesis of Compound 10

After 216 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 10 ml of anhydrous tetrahydrofuran, a solution of ethyl2-methylacetoacetate sodium salt separately prepared from 191 mg ofsodium hydride (60% content, oil dispersion) and 0.68 ml of ethyl2-methylacetoacetate in tetrahydrofuran was added to the solution at 0°C. The mixture was stirred at 20° C. for 20 minutes. The reactionmixture was poured into a phosphate buffer (pH 4) followed by extractionwith chloroform. The chloroform layer was washed with saturated aqueoussodium hydrogencarbonate solution, and dried over anhydrous sodiumsulfate, and then the solvent was distilled off. The residue obtainedwas purified by column chromatography (silica gel;chloroform:methanol=50:1) to give 160 mg (yield, 56%) ofIa-acetyl-6-[2-acetyl-2-(ethoxy-carbonylpropyl)]-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydromitomycinA.

The whole amount of the product obtained was dissolved in 20 ml ofmethanol and 8 mg of potassium carbonate was added to the solution. Themixture was stirred at 0° C. for 3 hours. The reaction mixture wasneutralized with a phosphate buffer (pH 6) followed by extraction withchloroform. The chloroform layer was dried over anhydrous sodium sulfateand the solvent was distilled off in vacuo. The residue obtained waspurified by preparative TLC (silica gel; chloroform:methanol=30:1 to20:1) to give 72 mg (yield, 50%) of the Ia-acetyl derivative of Compound10.

The whole amount of the product was dissolved in 8 ml of tetrahydrofuranand 1 ml of saturated aqueous sodium hydrogencarbonate solution. Themixture was stirred at ambient temperature for 4 hours. Saturatedaqueous sodium chloride solution was added to the reaction mixturefollowed by extraction with chloroform. The chloroform layer was driedover anhydrous sodium sulfate and the solvent was distilled off invacuo. The residue obtained was purified by preparative TLC (silica gel;chloroform-methanol=20:1) to give 20 mg (yield, 31%) of Compound 10 asthe mixture of diastereoisomers.

TLC: Rf=0.54 (chloroform:methanol=9:1)

SI-MS (m/z): 493 (M⁺ +2); C₂₃ H₂₉ N₃ O₉ =491

IR (cm⁻¹): 3430, 2930, 1710, 1620, 1570, 1440, 1330, 1280, 1210, 1110,1070

¹ H-NMR (400 MHz, chloroform -d₁) δ(ppm): Diastereoisomer A:1.1˜1.4 (br,1H), 1.19 (s, 3H), 1.29 (t, J=7.1 Hz, 3H), 2.22 (s, 3H), 2.81 (dd, J=1.7& 4.4 Hz, 1H), 2.90 (d, J=4.4 Hz, 1H), 2.98 (s, 2H), 3.22 (s, 3H), 3.45(bd, J=12.8 Hz, 1H), 3.60 (dd, J=4.4 & 10.6 Hz, 1H), 4.00 (d, J=12.6 Hz,1H), 4.06 (s, 3H), 4.15˜4.27 (m, 2H), 4.54 (t, J=10.6 Hz, 1H), 4.73 (dd,J=4.4 & 10.6 Hz, 1H), 4.90 (br, 2H)

Diastereoisomer B:1.1˜1.4 (br, 1H), 1.19 (s, 3H), 1.29 (t, J=7.1 Hz,3H), 2.21 (s, 3H), 2.81 (dd, J=1.7 & 4.4 Hz, 1H), 2.90 (d, J=4.4 Hz,1H), 2.98 (s, 2H), 3.22 (s, 3H), 3.45 (bd, J=12.8 Hz, 1H), 3.60 (dd,J=4.4 & 10.6 Hz, 1H), 3.97 (d, J=12.6 Hz, 1H), 4.06 (s, 3H), 4.15˜4.27(m, 2H), 4.54 (t, J=10.6 Hz, 1H), 4.71 (dd, J=4.4 & 10.8 Hz, 1H), 4.90(br, 2H)

EXAMPLE 11 Synthesis of Compound 11

After 196 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 10 ml of chloroform, a solution of diethyl methylmalonate sodium saltseparately prepared from 116 mg of sodium hydride (60% content, oildispersion) and 0.49 ml of diethyl methylmalonate in tetrahydrofuran wasadded to the solution at 0° C. The mixture was stirred at 20° C. for anhour. The reaction mixture was poured into a phosphate buffer (pH 4)followed by extraction with chloroform. The chloroform layer was washedwith saturated aqueous sodium hydrogencarbonate solution, and dried overanhydrous sodium sulfate, and then the solvent was distilled off. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give 160 mg (yield, 58%) ofla-acetyl-6-[2,2-bis(ethoxycarbonyl)propyl]-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxymitomycin A.

The whole amount of the product obtained was dissolved in 15 ml ofethanol and 1.0 ml of a 6.8M solution of ammonia in methanol was addedto the solution. The mixture was stirred at 20° C. for 10 hours. Thesolvent was distilled off in vacuo. The residue was purified bypreparative TLC (silica gel; chloroform:methanol=94:6) to give 44 mg(yield, 35%) of Compound 11.

TLC: Rf=0.51 (chloroform:methanol:acetone=4.5:0.5:5.0)

SI-MS(m/z): 463 (M⁺ +3); C₂₁ H₂₄ N₄ O₈ =460

IR (cm⁻¹): 3450, 2930, 1710, 1650, 1620, 1570, 1460, 1410, 1340, 1260,1160, 1100, 1070

¹ H-NMR (400 MHz, chloroform -d₁) δ (ppm): 0.90 (br, 1H), 1.23 (t, J=7.1Hz, 3H), 1.49 (s, 3H), 2.55 (d, J=17.5 Hz, 1H), 2.88 (bd, J=4.7 Hz, 1H),2.94 (d, J=4.7 Hz, 1H), 3.23 (s, 3H), 3.33 (d, J=17.5 Hz, 1H), 3.52 (dd,J=1.7 & 12.8 Hz, 1H), 3.66 (dd, J=4.7 & 10.3 Hz, 1H), 4.13 (d, J=12.8Hz, 1H), 4.20 (q, J=7.1 Hz, 2H), 4.56 (t, J=10.6 Hz, 1H), 4.67 (dd,J=4.7 & 10.6 Hz, 1H), 4.81 (br, 2H), 8.11 (br, 1H)

EXAMPLE 12 Synthesis of Compound 12

After 196 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 5 ml of chloroform, a solution of ethyl acetoacetate sodium saltseparately prepared from 70 mg of sodium hydride (60% content, oildispersion) and 0.22 ml of ethyl acetoacetate in tetrahydrofuran wasadded to the solution at 0° C. The mixture was stirred at 20° C. for 1.5hours. The reaction mixture was poured into a phosphate buffer (pH 4)followed by extraction with chloroform. The chloroform layer was washedwith saturated aqueous sodium hydrogencarbonate solution, and dried overanhydrous sodium sulfate, and then the solvent was distilled off. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=50:1) to give 120 mg (yield, 47%) of the la-acetylderivative of Compound 12.

150 mg of the product obtained was dissolved in 20 ml of methanol and300 mg of ammonium acetate was added to the solution. The reactionmixture was poured into saturated aqueous sodium chloride solutionfollowed by extraction with chloroform. The chloroform layer was driedover anhydrous sodium sulfate and the solvent was distilled off invacuo. The residue was purified by preparative TLC (silica gel;chloroform:methanol=30:1) to give 23.8 mg (yield, 20%) of Compound 12.

TLC: Rf=0.40 (chloroform:methanol=9:1)

SI-MS (m/z): 447 (M⁺ +2); C₂₁ H₂₃ N₃ O₈ =445

IR (cm⁻¹): 3450, 2910, 1720, 1650, 1630, 1570, 1440, 1390, 1360, 1340,1310, 1190, 1160, 1090, 1040

¹ H-NMR (400 MHz, chloroform -d₁) δ (ppm): 0.60 (br, 1H), 1.31 (t, J=7.1Hz, 3H), 2.38 (s, 3H), 2.85 (br, 1H), 2.91 (br, 1H), 3.09 (d, J=20.4 Hz,1H), 3.20 (d, J=20.4 Hz, 1H), 3.23 (s, 3H), 3.45 (bd, J=12.3 Hz, 1H),3.67 (dd, J=4.7 & 10.1 Hz, 1H), 4.07 (d, J=12.8 Hz, 1H), 4.23 (q, J=7.1Hz, 2H), 4.60 (bt, J=10.3 Hz, 1H), 4.70 (dd, J=4.7 & 10.6 Hz, 1H), 4.72(br, 2H)

EXAMPLE 13 Synthesis of Compound 13

1.03 g of Compound e obtained in REFERENCE EXAMPLE 5 was dissolved in 50ml of anhydrous tetrahydrofuran and 0.20 ml of anhydrous triethylamineand 346 mg of dimedone were added to the solution followed by stirringat 20° C. for 40 minutes. The reaction mixture was diluted withsaturated aqueous sodium chloride solution followed by extraction withchloroform. The organic layer was dried over anhydrous sodium sulfateand the solvent was distilled off. The residue was again dissolved in100 ml of chloroform and 100 ml of silica gel was added to the solutionto adsorb the product thereto. The reaction mixture was allowed to standat ambient temperature for 15 hours. Silica gel was extracted with aneluent (chloroform:methanol=9:1) and the resultant violet solution wasconcentrated in vacuo. The residue was purified by column chromatography(silica gel; chloroform:methanol=50:1) to obtain a violet fraction. Thisfraction was treated in a conventional manner to give 634.9 mg (yield,53%) of the la-allyloxycarbonyl derivative of Compound 13 as a violetpowder.

To a solution of 251.9 mg of the compound above in 20 ml oftetrahydrofuran were added 0.200 ml of triethylammonium formate and 30mg of tetrakis (triphenylphosphine) palladium (0) under argonatmosphere. The mixture was stirred at 20° C. for 22 minutes. Thereaction mixture was applied directly to column chromatography (silicagel; chloroform:methanol=20:1) to give a reddish violet fraction. Thisfraction was treated in a conventional manner to give 110.2 mg (yield,52%) of Compound 13 as a reddish violet powder.

TLC: Rf=0.29 (chloroform:methanol=9:1)

FAB-MS (m/z): 456 (M⁺ +1); C₂₃ H₂₅ N₃ O₇ =455

IR (cm⁻¹): 3450, 3300, 2950, 2880, 1720, 1710, 1660, 1630, 1570, 1380,1330, 1200, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ (ppm): 0.95 (s, 3H), 0.99 (s, 3H),2.12 (bs, 1H), 2.33 (bs, 4H), 2.78 (bs, 1H), 3.08 (d, J=20.5 Hz, 1H),3.16 (bs, 1H), 3.20 (d, J=20.7 Hz, 1H), 3.27 (s, 3H), 3.53(bd, J=12 Hz,1H), 4.06 (dd, J=4.4 & 11.0 Hz, 1H), 4.18 (d, J=12.5 Hz, 1H), 5.12 (bt,J=10 Hz, 1H), 5.42 (dd, J=4.3 & 10.4 Hz, 1H), 7.4˜7.8 (bs, 2H)

EXAMPLE 14 Synthesis of Compound 14

840 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolved in 50ml of anhydrous tetrahydrofuran and 0.20 ml of anhydrous triethylamineand 231.6 mg of 1,3-cyclohexanedione were added to the solution followedby stirring at 20° C. for 3 hours. The reaction mixture was diluted withsaturated aqueous sodium chloride solution followed by extraction withchloroform. After the organic layer was dried over anhydrous sodiumsulfate, the solvent was distilled off. The residue was purified bycolumn chromatography (silica gel; chloroform:methanol=20:1) to give aviolet fraction. This fraction was treated in a conventional manner togive 591.7 mg (yield, 63%) of the la-acetyl derivative of Compound 14 asa violet powder.

The product (552.1 mg) obtained above was dissolved in 50 ml of methanoland 323.5 mg of potassium carbonate was added to the solution followedby stirring at -20° C. for 1.25 hours. The reaction mixture was dilutedwith a phosphate buffer (pH 4), which was then extracted withchloroform. The organic layer was dried over anhydrous sodium sulfateand the solvent was distilled off. The residue was purified by columnchromatography (silica gel; chloroform:methanol=30:1) to give a reddishviolet fraction. This fraction was again purified by preparative TLC(silica gel; chloroform:methanol=9:1) and then treated in a conventionalmanner to give 70.4 mg (yield, 14%) of Compound 14 as a red violetpowder.

TLC: Rf=0.36 (chloroform:methanol=9:1)

FAB-MS (m/z): 429 (M⁺ +2); C₂₁ H₂₁ N₃ O₇ =427

IR (cm⁻¹): 3450, 3350, 3330, 3200, 2950, 2880, 1720, 1660, 1630, 1570,1450, 1380, 1340, 1210, 1190, 1120, 1070 ¹ H-NMR (270 MHz, pyridine -d₅)δ (ppm): 1.7˜1.9 (m, 2H), 2.13 (bs, 1H), 2.3˜2.5 (m, 4H), 2.79 (bs, 1H),3.10˜3.25 (m, 3H), 3.27 (s, 3H), 3.53 (bd, J=12 Hz, 1H), 4.04 (dd, J=4.3& 11.1 Hz, 1H), 4.18 (d, J=12.5 Hz, 1H), 5.09 (bt, J=11 Hz, 1H), 5.39(dd, J=4.3 & 10.4 Hz, 1H), 7.1˜7.8 (br, 2H)

EXAMPLE 15 Synthesis of 6-demethyl-6-ethylmitomycin A (Compound 15)

After 843 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 100 ml of anhydrous tetrahydrofuran, 1.94 g of phenol was added tothe solution. With stirring at -78° C., 40 ml of a 0.25M solution oflithium dimethyl cuprate in tetrahydrofuran was added dropwise to thesolution over 50 minutes. The mixture was stirred for 30 minutes at thesame temperature.

Saturated aqueous ammonium chloride solution was added to the yellowishorange reaction mixture. After the resulting reddish pink solution wasseparated, the aqueous layer was extracted with chloroform. The organiclayer was washed with saturated aqueous sodium chloride solution anddried over anhydrous sodium sulfate. The desiccant was removed byfiltration and the solvent was distilled off. The residue was purifiedby column chromatography (silica gel; chloroform:methanol=30:1 to 20:1)to give a yellowish pink fraction containing the desired product.n-Hexane-chloroform was added to this fraction to give a powder. Thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 382.6 mg (yield, 44%) ofla-acetyl-7-demethoxy-6-demethyl-6-ethyl-7-ethylenedioxy-6-,7-dihydromitomycin A as a yellowish pink powder.

371.9 mg of the product obtained above was dissolved in 40 ml ofmethanol and 237 mg of potassium carbonate was added to the solution.The mixture was stirred at ambient temperature for 2 hours and 50minutes.

Saturated aqueous sodium chloride solution was added to the reactionmixture and the mixture was extracted with chloroform. The organic layerwas washed with saturated aqueous sodium chloride solution and driedover anhydrous sodium sulfate. The desiccant was removed by filtrationand the solvent was distilled off. The residue was purified by columnchromatography (silica gel; chloroform:acetone=2:1) to give a violetfraction containing the desired product. After the solvent was distilledoff from this fraction, n-hexane-chloroform was added to give a powder.The solvent was distilled off and the residue was thoroughly dried invacuo to give 187.1 mg (yield, 60%) of Compound 15 as a violet powder.

TLC: Rf=0.32 (chloroform:acetonitrile:n-hexane=5:5:1)

FAB-MS (m/z): 365 (M⁺ +2); C₁₇ H₂₁ N₃ O₆ =363

IR (cm⁻¹): 3450, 3300, 2950, 1740, 1710, 1650, 1630, 1570, 1330, 1270,1220, 1050

¹ H-NMR (270 MHz, pyridine -d₅) δ (ppm): 0.98 (t, J=7.2 Hz, 3H), 2.15(bt, J=7.4 Hz, 1H), 2.40 (q, J=7.2 Hz, 2H), 2.78 (bs, 1H), 3.15 (dd,J=4.5 & 7.4 Hz, 1H), 3.23 (s, 3H), 3.54 (bd, J=ca. 12 Hz, 1H), 4.01 (dd,J=4.2 & 10.9 Hz, 1H), 4.04 (s, 3H), 4.24 (d, J=12.4 Hz, 1H), 5.09 (bt,J=ca. 11 Hz, 1H), 5.42 (dd, J=4.2 & 10.1 Hz, 1H), 7.3˜8.0 (br, 2H)

EXAMPLE 16 Synthesis of 6-demethyl-6-ethylmitomycin B (Compound 16)

After 375 mg of7-demethoxy-6-demethyl-6,7-dihydro-6-ethyl-7-ethylenedioxymitomycin Bobtained in EXAMPLE 3 was dissolved in solution followed by stirring atambient temperature for 4 hours and 10 minutes.

The reaction mixture was subjected to the same work-up as in EXAMPLE 15.The residue obtained was purified by column chromatography (silica gel;chloroform:methanol=20:1) to give a violet fraction containing thedesired product. After the solvent was distilled off from this fraction,n-hexane-chloroform was added to the residue to give a powder. Thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 143.6 mg (yield, 41%) of Compound 16 as a violet powder.

TLC: Rf=0.33 (chloroform:methanol=9:1)

FAB-MS (m/z): 366 (M⁺ +3); C₁₇ H₂₁ N₃ O₆ =363

IR (cm⁻¹): 3440, 3300, 3200, 2950, 1740, 1700, 1650, 1620, 1570, 1460,1450, 1340, 1260, 1110, 1060, 1040

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 0.92 (t, J=7.7 Hz, 3H), 2.11 (s,3H), 2.24 (dd, J=2.0 & 4.5 Hz, 1H), 2.3˜2.4 (m, 2H), 2.48 (d, J=4.5 Hz,1H), 3.60 (dd, J=2.0 & 12.9 Hz, 1H), 4.00 (s, 3H), 4.16 (d, J=12.4 Hz,1H), 4.24 (dd, J=3.5 & 9.9 Hz, 1H), 5.21 (t, J=10.1 Hz, 1H), 5.46 (dd,J=3.5 & 10.4 Hz, 1H), 7.3˜7.7 (br, 2H), 8.28 (s, 1H)

EXAMPLE 17 Synthesis of 6-demethyl-6-(2-methylpropyl)mitomycin C(Compound 17)

To a solution of 907 mg of Compound e obtained in REFERENCE EXAMPLE 5 in30 ml of anhydrous tetrahydrofuran, was added 113 mg of copper (I)iodide. The mixture was cooled to -78° C. While stirring, 5.0 ml of a0.67M solution of isopropylmagnesium bromide in tetrahydrofuran wasadded dropwise to the solution over an hour. After reaction mixture wasstirred at -78° C. for 20 minutes. 1.5 ml of the solution of isopropylmagnesium bromide in tetrahydrofuran was additionally added dropwise tothe reaction mixture over 25 minutes and the resulting mixture wasstirred at -78° C. for 5 minutes.

Saturated aqueous ammonium chloride solution was added to the yellowishorange reaction mixture. After the resultant reddish pink solution wasseparated, the aqueous layer was extracted with chloroform. The organiclayer was washed with saturated aqueous sodium chloride solution, anddried over anhydrous sodium sulfate. The desiccant was removed byfiltration, and the solvent was distilled off. The residue obtained waspurified by column chromatography (silica gel; chloroform:methanol=20:1)to give a yellowish pink fraction containing the desired product.n-Hexane-chloroform was added to the mixture to give a powder. Thesolvent was distilled off and then thoroughly dried in vacuo to give587.2 mg (yield, 59%) ofIa-allyloxycarbonyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-(2-methylpropyl)mitomycinA as a yellowish pink powder.

259.6 mg of the product obtained above was dissolved in 30 ml ofanhydrous tetrahydrofuran. The solution was allowed to stand for 137.5hours at ambient temperature under anhydrous ammonia atmosphere.

The reaction mixture was concentrated under reduced pressure and theresidue obtained was purified by preparative TLC (silicagel:chloroform:methanol=9:1) to give a violet fraction containing thedesired product. This fraction was extracted with the developing solventand the solvent was distilled off. Then, n-hexane-chloroform was addedto the residue to give a powder. After stripping the solvent, theresidue was thoroughly dried in vacuo to give 163.2 mg (yield, 69%) ofIa-allyloxycarbonyl derivative of Compound 17 as a violet powder.

191.8 mg of the product obtained above was dissolved in 20 ml ofanhydrous tetrahydrofuran, and 0.20 ml of triethylammonium formate and asolution of 30 mg of tetrakis (triphenylphosphine) palladium (0) in 2.0ml of anhydrous tetrahydrofuran was added to the solution. The mixturewas stirred at ambient temperature for an hour and 10 minutes under anargon atmosphere.

The reaction mixture was purified directly by column chromatography(silica gel; chloroform:methanol=30:1 to 20:1). The solvent wasdistilled off from the resulting violet fraction. Then,n-hexane-chloroform was added to give a powder. After stripping solvent,the residue was thoroughly dried in vacuo to give 112.2 mg (yield, 72%)of compound 17 as a violet powder.

TLC: Rf=0.29 (chloroform:methanol=9:1)

FAB-MS (m/z): 377 (M⁺ +1); C₁₈ H₂₄ N₄ O₅ =376

IR (cm⁻¹): 3480, 3440, 3430, 3330, 3280, 2950, 1730, 1600, 1560, 1450,1370, 1330, 1070

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 0.91 (d, J=6.4 Hz, 6H), 1.95˜2.16(m, 2H), 2.51 (d, J=7.4 Hz, 2H), 2.74 (bs, 1H), 3.15 (bs, 1H), 3.21 (s,3H), 3.61 (bd, J=ca. 13 Hz, 1H), 4.05 (dd, J=4.5 & 10.9 Hz, 1H), 4.58(d, J=12.9 Hz, 1H), 5.13 (bt, J=ca. 10 Hz, 1H), 5.45 (dd, J=10.4 Hz,1H), 7.3˜7.9 (br, 4H)

EXAMPLE 18 Synthesis of 6-demethyl-6-(2-methylpropyl)mitomycin A(Compound 18)

After 400.2 mg ofIa-allyloxycarbonyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-(2-methylpropyl)mitomycin A was dissolved in 25 ml of anhydrous tetrahydrofuran, 1.0 mlof triethylammonium formate and a solution of 94 mg of tetrakis(triphenylphosphine) palladium (0) in 4.5 ml of tetrahydrofuran wereadded to the solution in 3 portions, respectively. The mixture wasstirred for 3 hours at ambient temperature under an argon atmosphere.

The reaction mixture was applied directly to column chromatography(silica gel; chloroform:methanol=50:1 to 20:1) to give a fractioncontaining the desired product. The solvent was distilled off from thisfraction. Chloroform-n-hexane was added to the residue to give a powder.The solvent was distilled off and the residue was thoroughly dried invacuo to give 247 mg of crude7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-(2-methylpropyl)mitomycinA as a yellowish pink powder.

The crude product (223.8 mg, 90.6% of the whole amount) obtained abovewas dissolved in 30 ml of methanol and 148 mg of potassium carbonate wasadded to the solution followed by stirring at ambient temperature for anhour.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by column chromatography (silica gel;chloroform:acetonitrile:n-hexane=5:5:1) to give a reddish violetfraction containing the desired product. After the solvent was distilledoff from this fraction, chloroform-n-hexane was added to give a powder.The solvent was distilled off and the residue was thoroughly dried invacuo to give 36.7 mg (yield, 18%) of Compound 18 as a reddish as areddish violet powder.

TLC: Rf=0.44 (chloroform:methanol=9:1)

FAB-MS (m/z): 392 (M⁺ +1); C₁₉ H₂₅ N₃ O₆ =391

IR (cm⁻¹): 3450, 3350, 3300, 2950, 1720, 1710, 1650, 1630, 1570, 1460,1450, 1410, 1340, 1310, 1240, 1080, 1070, 1040

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 0.86 (d, J=6.4 Hz, 6H), 1.85(m,1H), 2.12 (m, 1H), 2.35(m, 2H), 2.77(bs, 1H), 3.15(bs, 1H), 3.22 (s,3H), 3.55 (bd, J=12.4 Hz, 1H), 4.01 (dd, J=4.5 & 11.4 Hz, 1H), 4.07 (s,3H), 4.25 (bd, J=12.4 Hz, 1H), 5.08 (bt, J=10.6 Hz, 1H), 5.42 (dd, J=4.5& 10.4 Hz, 1H), 7.4˜7.9(br, 2H)

EXAMPLE 19 Synthesis of 6-demethyl-6-(2-methylpropyl) mitomycin B(Compound 19)

After 645 mg of Compound c obtained in REFERENCE EXAMPLE 3 was dissolvedin 50 ml of anhydrous tetrahydrofuran, with stirring at -78° C., 3.0 mlof a 0.67M solution of isopropyl magnesium bromide in tetrahydrofuranwas added dropwise to the solution over 15 minutes. The mixture wasstirred for an hour and 40 minutes under the same conditions.

The reaction mixture was subjected to work-up as in EXAMPLE 17. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1 to 20:1) to give a yellowish pink fractioncontaining the desired product. Chloroform-n-hexane was added to thefraction to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 171 mg (yield, 30% based on7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin B) of7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-(2-methylpropyl)mitomycinB as a yellowish pink powder.

After 161.4 mg of the product obtained above was dissolved in 30 ml ofmethanol, 110 mg of potassium carbonate was added to the solutionfollowed by stirring at ambient temperature for 55 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by preparative TLC (silica gel;chloroform:methanol=9:1) to give a violet fraction containing thedesired product. After the solvent was distilled off from this fraction,chloroform-n-hexane was added to give a powder. The solvent wasdistilled off and the residue was thoroughly dried in vacuo to give 63.3mg (yield, 42%) of Compound 19 as a violet powder.

TLC: Rf=0.44 (chloroform:methanol=9:1)

FAB-MS (m/z): 393 (M⁺ +2); C₁₉ H₂₅ N₃ O₆ =391

IR (cm⁻¹): 3430, 3350, 3300, 2950, 1730, 1690, 1650, 1620, 1610, 1560,1460, 1440, 1330, 1310, 1360, 1240, 1110

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 0.82 (d, J=6.9 Hz, 6H), 1.78 (m,1H), 2.10 (s, 3H), 2.24 (dd, J=2.0 & 5.0 Hz, 1H), 2.28 (m, 2H), 2.47 (d,J=5.0 Hz, 1H), 3.61 (dd, J=2.0 & 12.4 Hz, 1H), 4.02(s, 3H), 4.16(d,J=12.9 Hz, 1H), 4.25 (dd, J=3.5 & 9.4 Hz, 1H), 5.20 (bt, J=10.2 Hz, 1H),5.44 (dd, J=3.5 & 10.9 Hz, 1H), 7.3˜7.8 (br, 2H), 8.28 (s, 1H)

EXAMPLE 20 Synthesis of 6-demethyl-6-n-pentylmitomycin A (Compound 20)

After 1.158 g of Compound e obtained in REFERENCE EXAMPLE 5 wasdissolved in 80 ml of anhydrous tetrahydrofuran, 40 ml of a solution oflithium di-n-butyl cuprate in tetrahydrofuran-n-hexane (about 5.0mmoles) was added dropwise to the solution over 30 minutes whilestirring at -78° C. Subsequently, the mixture was stirred at -78° C. to-20° C. for 50 minutes.

The yellowish orange reaction mixture was subjected to work-up as inEXAMPLE 17. The residue obtained was purified by column chromatography(silica gel; chloroform:methanol=30:1) to give a yellowish pink fractioncontaining the desired product. Chloroform-n-hexane was added to thisfraction to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 472.9 mg (yield, 43%) of7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-pentylmitomycin Aas a yellowish pink powder.

After 403.2 mg of the product obtained above was dissolved in 50 ml ofmethanol, 511 mg of sodium hydrogen carbonate was added to the solutionfollowed by stirring at ambient temperature for 23 hours and 20 minutes.

Chloroform was added to the reaction mixture and the precipitatedinorganic salt was filtered off. After the solvent was distilled off,the residue was purified by column chromatography (silica gel;chloroform:methanol=50:1) to give a reddish violet fraction containingthe desired product. After the solvent was distilled off from thisfraction, chloroform-n-hexane was added to give a powder. The solventwas distilled off and the residue was thoroughly dried in vacuo to give264.5 mg (yield, 70%) of Compound 20 as a reddish violet powder.

TLC: Rf=0.37 (chloroform:methanol=9:1)

FAB-MS (m/z): 406 (M⁺ +1); C₂₀ H₂₇ N₃ O₆ =405

IR (cm⁻¹): 3400, 3300, 2950, 2900, 1740, 1700, 1650, 1630, 1580, 1570,1450, 1430, 1410, 1340, 1320, 1270, 1220, 1070.

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.62 (bs, 1H), 0.89 (bt, J=6.9Hz, 3H), 1.23˜1.43 (m, 6H), 2.22˜2.41 (m, 2H), 2.84 (bs, 1H), 2.91 (bs,1H), 3.23 (s, 3H), 3.48 (bd, J=12.9 Hz, 1H), 3.62 (dd, J=4.6 & 10.6 Hz,1H), 4.04 (s, 3H), 4.05 (d, J=12.9 Hz, 1H), 4.58 (bt, J=10.4 Hz, 1H),4.74 (dd, J=4.6 & 10.4 Hz, 1H), 4.7˜4.8 (br, 2H)

EXAMPLE 21 Synthesis of 6-demethyl-6-n-pentylmitomycin B (Compound 21)

After 642 mg of Compound c obtained in REFERENCE EXAMPLE 3 was dissolvedin 50 ml of anhydrous tetrahydrofuran, 2.0 g of phenol was added to thesolution. While stirring at -78° C., 41 ml of a solution of lithiumdi-n-butyl cuprate in tetrahydrofuran-n-hexane (about 0.31M) was addeddropwise to the solution over 15 minutes. Subsequently, the mixture wasstirred for 25 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 17. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1 to 10:1) to give a yellowish pink fractioncontaining the desired product. n-Hexane-chloroform was added to thisfraction to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 174 mg (yield, 30% based on7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin B) of7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-pentylmitomycin Bas a yellowish pink powder.

After 164.9 mg of the product obtained above was dissolved in 30 ml ofmethanol, 108 mg of potassium carbonate was added to the solutionfollowed by stirring at ambient temperature for 50 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by preparative TLC (silica gel;chloroform:acetone=1:1) to give a violet fraction containing the desiredproduct. This fraction was extracted with the developing solvent and thesolvent was distilled off. Then, chloroform-n-hexane was added to give apowder, the solvent was distilled off and the residue was thoroughlydried in vacuo to give 96.1 mg (yield, 63%) of Compound 21 as a violetpowder.

TLC: Rf=0.51 (chloroform:methanol=9:1)

FAB-MS (m/z): 407 (M⁺ +2); C₂₀ H₂₇ N₃ O₆ =405

IR (cm⁻¹): 3430, 3350, 3280, 2950, 2930, 2850, 1730, 1690, 1650, 1620,1570, 1460, 1450, 1330, 1260, 1210, 1120

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 0.80 (bt, J=6.9 Hz, 3H), 1.1˜1.4(m, 6H), 2.11 (s, 3H), 2.23 (dd, J=1.7 & 4.7 Hz, 1H), 2.3˜2.4 (m, 2H),2.47 (d, J=4.5 Hz, 1H), 3.60 (dd, J=2.0 & 12.9 Hz, 1H), 4.03 (s, 3H),4.17 (d, J=12.9 Hz, 1H), 4.24 (dd, J=3.5 & 9.4 Hz, 1H), 5.20 (bt, J=10.2Hz, 1H), 5.44 (dd, J=3.5 & 10.9 Hz, 1H), 7.3˜7.7 (br, 2H), 8.24 (s, 1H)

EXAMPLE 22 Synthesis of 6-benzyl-6-demethylmitomycin A (Compound 22)

After 845 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 100 ml of anhydrous tetrahydrofuran, 1.95 g of phenol was added tothe solution. While stirring at -78° C., 30 ml of a solution of lithiumdiphenyl cuprate in tetrahydrofuran (about 0.34M) was added dropwise tothe solution over 15 minutes. Subsequently, the mixture was stirred for50 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 17. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give a yellowish pink fraction containingthe desired product. n-Hexane-chloroform was added to give a powder. Thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 584.1 mg (yield, 58%) ofla-acetyl-6-benzyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydromitomycinA as a yellowish pink powder.

After 574.7 mg of the product obtained above was dissolved in 50 ml ofmethanol, 320 mg of potassium carbonate was added to the solutionfollowed by stirring at ambient temperature for 1.5 hours.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol,=50:1 to 30:1) to give a reddish violet fractioncontaining the desired product. After the solvent was distilled off fromthis fraction, chloroform-n-hexane was added to give a powder, thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 238.4 mg (yield, 49%) of Compound 22 as a reddish violet powder.

TLC: Rf=0.43 (chloroform:methanol=9:1)

SI-MS (m/z): 426 (M⁺ +1); C₂₂ H₂₃ N₃ O₆ =425

IR (cm⁻¹): 3450, 3370, 3300, 2950, 1720, 1710, 1630, 1570, 1450, 1440,1340, 1330, 1320, 1220, 1070, 1050

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 2.12 (bs, 1H), 2.74 (bs, 1H),3.11 (bs, 1H), 3.17 (s, 3H), 3.52 (bd, J=12.9 Hz, 1H), 3.75 (d, J=13.7Hz, 1H), 3.81 (d, J=13.7 Hz, 1H), 3.97 (dd, J=4.2 & 11.2 Hz, 1H), 4.07(s, 3H), 4.21 (d, J=12.9 Hz, 1H), 5.04 (bt, J=ca. 11 Hz, 1H), 5.37 (dd,J=4.2 & 10.6 Hz, 1H), 7.15˜7.35 (m, 3H), 7.40˜7.45 (m, 2H), 7.3˜7.9 (br,2H)

EXAMPLE 23 Synthesis of 6-demethyl-6-benzylmitomycin B (Compound 23)

After 963 mg of Compound c obtained in REFERENCE EXAMPLE 3 was dissolvedin 100 ml of anhydrous tetrahydrofuran, 2.26 g of phenol was added tothe solution. While stirring at -78° C., 40 ml of lithium diphenylcuprate in tetrahydrofuran (about 0.34M) was added dropwise to thesolution over 20 minutes. Subsequently, the mixture was stirred for 50minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 17. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=20:1) to give a yellowish pink fraction containingthe desired product. n-Hexane-chloroform was added to give a powder. Thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 208.2 mg (yield, 21% based on7-demethoxy-7-ehylenedioxy-6,7-dihydro-6-phenylselenomitomycin B) of6-benzyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydromitomycin Bas a yellowish pink powder.

After 194.5 mg of the product obtained above was dissolved in 30 ml ofmethanol, 128 mg of potassium carbonate was added to the solutionfollowed by stirring at ambient temperature for an hour and 10 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give a violet fraction containing thedesired product. After the solvent was distilled off from this fraction,chloroform-n-hexane was added to give a powder. The solvent wasdistilled off and the residue was thoroughly dried in vacuo to give148.3 mg (yield, 82%) of Compound 23 as a violet powder.

TLC: Rf=0.33 (chloroform:methanol=9:1)

FAB-MS (m/z): 427 (M⁺ +2); C₂₂ H₂₃ N₃ O₆ =425

IR (cm⁻¹): 3400, 3330, 2900, 1750, 1740, 1660, 1600, 1490, 1450, 1370,1310, 1240, 1110

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 2.09 (s, 3H), 222 (dd, J=1.6 &4.5 Hz, 1H), 2.44 (d, J=4.5 Hz, 1H), 3.57 (dd, J=1.6 & 12.8 Hz, 1H),3.71 (s, 2H), 4.01 (s, 3H), 4.10 (d, J=12.8 Hz, 1H), 4.21 (dd, J=3.5 &9.9 Hz, 1H), 5.15 (t, J=10.1 Hz, 1H), 5.40 (dd, J=3.5 & 10.4 Hz, 1H),7.10˜7.36 (m, 5H), 7.48 (bs, 2H), 8.26 (s, 1H)

EXAMPLE 24 Synthesis of 6-demethyl-6-n-undecylmitomycin C (Compound 24)

After 808 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 20 ml of anhydrous tetrahydrofuran, 72 mg of copper (I) iodide wasadded to the solution. The mixture was cooled to -20° C. While stirringat this temperature, 6.0 ml of a 0.097M solution of n-decylmagnesiumbromide in tetrahydrofuran was added dropwise to the solution over 50minutes. Subsequently, the mixture was stirred for 30 minutes at -20° C.to 0° C.

The yellowish orange reaction mixture was subjected to work-up as inEXAMPLE 17. The residue obtained was purified by column chromatography(silica gel; chloroform:methanol=50:1) to give a yellowish pink fractioncontaining the desired product. n-Hexane-chloroform was added to thisfraction to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 131.6 mg (yield, 12%) ofla-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-undecylmitomycinA as yellowish pink paste.

The whole amount of the product obtained above was dissolved in 20 ml ofmethanol and 5 ml of a 6.8M solution of ammonia in methanol and thesolution was allowed to stand at ambient temperature for 17 hours and 40minutes.

The reaction mixture was concentrated under reduced pressure. Theresidue obtained was purified by preparative TLC (silicagel:chloroform:methanol=9:1) to give a violet fraction containing thedesired product. This fraction was extracted with the developingsolvent. After the solvent was distilled off, n-hexane-chloroform wasadded to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 9.1 mg (yield, 8%) of Compound 24as a violet powder.

TLC: Rf=0.31 (chloroform:methanol=9:1)

FAB-MS (m/z): 476 (M⁺ +2), 515[(M⁺ +2)+K⁺ ]: C₂₅ H₃₈ N₄ O₅ =474

IR (cm⁻¹): 3450, 3330, 3200, 2920, 2850, 1720, 1710, 1600, 1570, 1550,1450, 1360, 1340, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 0.86 (t, J=6.6 Hz, 3H), 1.1˜1.4(m, 16H), 1.56 (m, 2H), 2.0˜2.3 (br, 1H), 2.63 (m, 2H), 2.75 (bs, 1H),3.15 (bs, 1H), 3.22 (s, 3H), 3.62 (bd, J=12.7 Hz, 1H), 4.06 (dd, J=4.2 &11.2 Hz, 1H), 4.60 (d, J=12.7 Hz, 1H), 5.10 (bt, J=ca. 10 Hz, 1H), 5.46(dd, J=4.2 & 10.3 Hz, 1H), 7.4˜7.9 (br, 4H)

EXAMPLE 25 Synthesis of 6-demethyl-6-n-undecylmitomycin A (Compound 25)

To a solution of 848 mg of Compound a obtained in REFERENCE EXAMPLE 1 in100 ml of anhydrous tetrahydrofuran, was added 81 mg of copper (I)iodide. The mixture was cooled to -20° C. While stirring at -20° C., 7.0ml of a 0.86M solution of n-decylmagnesium bromide in tetrahydrofuranwas added dropwise to the mixture over 30 minutes. Subsequently, themixture was stirred for 20 minutes at -20° C.

The yellowish orange reaction mixture was subjected to work-up as inEXAMPLE 17. The residue obtained was purified by column chromatography(silica gel; chloroform:methanol=40:1 to 20:1) to give a yellowish pinkfraction containing the desired product. The solvent was distilled offfrom this fraction and the residue was thoroughly dried in vacuo to give495.1 mg ofla-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-undecylmitomycinA as yellowish pink paste.

The whole amount of the product obtained above was dissolved in 80 ml ofmethanol and 213 mg of potassium carbonate was added to the solutionfollowed by stirring at ambient temperature for 4 hours.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give a reddish violet fraction containingthe desired product. This fraction was extracted with the developingsolvent. After the solvent was distilled off, n-hexane-chloroform wasadded to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 159.3 mg (yield, 16% based onCompound a) of Compound 25 as a reddish violet powder.

TLC: Rf=0.57 (chloroform:methanol=9:1)

FAB-MS (m/z): 490 (M⁺ +1); C₂₆ H₃₉ N₃ O₆ =489

IR (cm⁻¹): 3450, 3350, 3300, 3200, 2920, 2850, 1720, 1650, 1630, 1580,1450, 1340, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ(ppm): 0.87 (bt, J=6.7 Hz, 3H), 1.0˜1.4(br, 16H), 1.43 (m, 2H), 2.0˜2.2 (br, 1H), 2.45 (m, 2H), 2.76 (bs, 1H),3.10 (bs, 1H), 3.22 (s, 3H), 3.54 (bd, J=ca,. 12 Hz, 1H), 3.96 (dd,J=4.4 & 11.1 Hz, 1H), 4.08 (s, 3H), 4.24 (d, J=12.4 Hz, 1H), 4.99 (bt,J=ca. 11 Hz, 1H), 5.34 (dd, J=4.4 & 10.4 Hz, 1H), 7.2˜7.9 (br, 2H)

EXAMPLE 26 Synthesis of 6-demethyl-6-n-nonadecylmitomycin C (Compound26)

To a solution of 1,265 g of Compound a obtained in REFERENCE EXAMPLE 1in 30 ml of anhydrous tetrahydrofuran, was added 96 mg of copper (I)iodide. The mixture was cooled to -78° C. While stirring -78° C. 8.0 mlof a 0.086M solution of octadecylmagnesium bromide in tetrahydrofuranwas added dropwise to the mixture over 25 minutes. Subsequently, themixture was stirred for 35 minutes at -78° C., and was stirred forfurther 40 minutes while elevating to ambient temperature.

The yellowish orange reaction mixture was subjected to work-up as inEXAMPLE 17. The residue obtained was purified by column chromatography(silica gel; chloroform:methanol=50:1) to give a yellowish pink fractioncontaining the desired product. n-Hexane-chloroform was added to give apowder. The solvent was distilled off and the residue was thoroughlydried in vacuo to give 404.6 mg (yield, 20%) ofla-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-n-nonadecylmitomycinA as a yellowish pink powder.

After 200.2 mg of the product obtained above was dissolved in the mixedsolution of 20 ml of methanol and 5 ml of a 6.8M solution of ammonia inmethanol. The resulting solution was allowed to stand at ambienttemperature for 20 hours and 30 minutes.

The reaction mixture was concentrated under reduced pressure and theresidue obtained was purified by column chromatography (silicagel:chloroform:methanol=40:1 to 30:1) and then by preparative TLC(silica gel; chloroform:methanol=9:1) to give a violet fractioncontaining the desired product. This fraction was extracted with thedeveloping solvent and the solvent was distilled off. Then,n-hexane-chloroform was added to the mixture to give a powder. Thesolvent was distilled off and then the residue was thoroughly dried invacuo to give 8.6 mg (yield, 5%) of Compound 26 as a violet powder.

TLC: Rf=0.38 (chloroform:methanol=9.1)

FAB-MS(m/z): 587 (M⁺ +1); C₃₃ H₅₄ N₄ O₅ =586

IR (cm⁻¹): 3450, 3330, 3200, 2920, 2850, 1720, 1600, 1570, 1560, 1450,1340, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ (ppm): 0.87 (t, J=6.5 Hz, 3H), 1.1˜1.5(m, 32H), 1.56 (m, 2H), 2.0˜2.2 (br, 1H), 2.63 (m, 2H), 2.74 (bs, 1H),3.14 (bs, 1H), 3.22 (s, 3H), 3.61 (bd, J=ca. 13 Hz, 1H), 4.05 (dd, J=4.2& 11.2 Hz, 1H), 4.60 (d, J=12.7 Hz, 1H), 5.11 (bt, J=ca. 10 Hz, 1H),5.45(dd, J=4.3 & 10.4 Hz, 1H), 7.4˜7.8 (br, 4H)

EXAMPLE 27 Synthesis of 6-demethyl-6-n-nonadecylmitomycin A (Compound27)

After 186.8 mg ofla-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-nonadecylmitomycinA was dissolved in 20 ml of methanol, 138 mg of potassium carbonate wasadded to the solution followed by stirring at ambient temperature for 6hours and 40 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 15. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=50:1) and then by preparative TLC (silicagel:chloroform:methanol=9:1) to give a reddish violet fractioncontaining the desired product. This fraction was extracted with thedeveloping solvent and the solvent was distilled off. Then,n-hexane-chloroform was added to give a powder. The solvent wasdistilled off and then the residue was thoroughly dried in vacuo to give4.9 mg (yield, 3%) of Compound 28 as a reddish violet powder.

TLC: Rf=0.55 (chloroform:methanol=9:1)

FAB-MS (m/z): 602 (M⁺ +1); C₃₄ H₅₅ N₃ O₆ =601

IR (cm⁻¹): 3450, 3420, 2920, 2850, 1710, 1630, 1570, 1470, 1450, 1340,1220, 1070

¹ H-NMR (270 MHz, pyridine -d₅) δ (ppm): 0.87 (t, J=6.7 Hz, 3H), 1.2˜1.4(m, 32H), 1.4˜1.5(m, 2H), 2.13 (bs, 1H), 2.47(bt, J=6.6 Hz, 2H), 2.77(bs, 1H), 3.15(bs, 1H), 3.24(s, 3H), 3.57(bd, J=12.5 Hz, 1H), 4.02 (dd,J=4.3 & 10.9 Hz, 1H), 4.10 (s, 3H), 4.28 (d, J=12.3 Hz, 1H), 5.09 (bt,J=ca. 10 Hz, 1H), 5.43 (dd, J=4.4 & 10.5 Hz, 1H), 7.5˜7.9(br, 2H)

EXAMPLE 28 Synthesis of Compound 28 and Compound 33

392 mg of Compound f obtained in REFERENCE EXAMPLE 6 was dissolved in 30ml of anhydrous tetrahydrofuran. While adding a suspension of the sodiumsalt prepared from 239 mg of ethyl p-nitrobenzoylacetate and 50.0 mg ofsodium hydride (60% content, oil dispersion) in 10 ml of tetrahydrofuranin several portions, the solution was stirred at ambient temperature foran hour and 50 minutes.

The reaction mixture was diluted with a phosphate buffer (pH 4) followedby extracting with chloroform. After washing with saturated aqueoussodium chloride solution, the organic layer was dried over anhydroussodium sulfate and the desiccant was removed by filtration. The solventwas then distilled off. The residue obtained was purified by columnchromatography (silica gel; chloroform:methanol=30:1) to give violet andyellow fractions, respectively. After the solvent was distilled off fromthese fractions, n-hexane-chloroform was added to give a power. Thesolvent was distilled off and the residue was thoroughly dried in vacuoto give 45.7 mg (yield, 8%) of Compound 33 as a violet powder.

The yellow fraction (20 mg) was dissolved in 5 ml of methanol and 200 mgof silica gel was added to the solution followed by refluxing for 4hours and 10 minutes with stirring. The reaction mixture was treated asdescribed above to give 10.2 mg (yield, 12%) of Compound 28 as a reddishviolet powder. Compound 28 was obtained as the mixture ofdiastereoisomers associated with the side chain of the 6-position methyland the proportion was approximately 1:1 by ¹ H-NMR.

Compound 33

TLC: Rf=0.64 (chloroform:methanol=9:1)

FAB-MS (m/z): 569 (M⁺ +3); C₂₇ H₂₆ N₄ O₁₀ =566

IR (cm⁻¹): 3400, 3340, 2900, 1760, 1740, 1690, 1660, 1610, 1550, 1380,1240, 1090

¹ H-NMR (270 MHz, chloroform-d₁) δ (ppm): 1.07 (t, J=7.2 Hz, 3H), 2.28(s, 3H), 2.2-2.4 (2H, overlapped with the peak of laN-methyl), 3.22 (s,3H), 3.33 (d, J=21.1 Hz, 1H), 3.42 (d, J=21.1 Hz, 1H), 3.47 (dd, J=1.7and 12.6 Hz, 1H), 3.65 (dd, J=4.7 & 10.6 Hz, 1H), 4.06 (d, J=ca. 13 Hz,1H, overlapped with the peak of ethyl), 4.07 (q, J=7.2 Hz, 2H), 4.40 (t,J=10.6 Hz, 1H), 4.70 (dd, J=4.7 & 10.6 Hz, 1H), 4.72 (bs, 2H), 7.59 (d,J=8.9 Hz, 2H), 8.25 (d, J=8.9 Hz, 2H)

Compound 28

TLC: Rf=0.64 (chloroform:methanol=9:1)

FAB-MS (m/z): 601 (M⁺ +3); C₂₈ H₃₀ N₄ O₁₁ =598

IR (cm⁻¹): 3400, 3350, 2900, 1760, 1740, 1660, 1560, 1480, 1370, 1320,1250, 1100

¹ H-NMR (270 MHz, chloroform-d₁) δ (ppm): 1.13 and 1.14 (t, J=6.9 Hz,3H), 2.27 and 2.28 (s, 3H), 2.2-2.3 (2H, overlapped with the peak of1aN-methyl), 2.94 and 2.98 (dd, J=5.9 & 13.9 Hz, 1H), 3.2-3.3 (m, 1H,6-CH₂, overlapped with the peak of 9a-methoxy), 3.18 and 3.20 (s, 3H),3.43 (dd, J=1.7 & 12.5 Hz, 1H), 3.55 and 3.57 (dd, J=4.5 & 10.9 Hz, 1H),4.00 (d, J=12.5 Hz, 1H), 4.09 (s, 3H), 4.0-4.2 (m, 2H), 4.33 and 4.34(t, J=10.6 Hz, 1H), 4.49-4.57 (m, 1H), 4.70 (dd, J= 4.5 & 10.4 Hz, 1H),4.76 (bs, 2H), 8.12 and 8.13 (d, J=8.9 Hz, 2H), 8.31 (d, J=8.9 Hz, 2H)

EXAMPLE 29 Synthesis of Compound 29

405 mg of Compound f obtained in REFERENCE EXAMPLE 6 was dissolved in 30ml of anhydrous tetrahydrofuran. While adding a suspension of the sodiumsalt prepared from 110 μl of acetylacetone and 52.5 mg of sodium hydride(60% content, oil dispersion) in 10 ml of tetrahydrofuran in severalportions, the solution was stirred at ambient temperature for 30minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=50:1) to give a violet fraction containing thedesired product. After the solvent was distilled off from this fraction,n-hexane-chloroform was added to give a powder. The solvent wasdistilled off and the residue was thoroughly dried in vacuo to give 31.8mg (yield, 6%) of Compound 29 as a violet powder.

TLC: Rf=0.38 (chloroform:methanol=9:1)

FAB-MS (m/z): 430 (M⁺ +1), 432 (M⁺ +3); C₂₁ H₂₃ N₃ O₇ =429

IR (cm⁻¹): 3450, 3350, 2950, 2920, 1730, 1710, 1690, 1660, 1630, 1570,1450, 1350, 1320, 1200, 1190, 1080

¹ H-NMR (270 MHz, chloroform -d₁) δ (ppm): 2.27 (s, 3H), 2.30 (s, 3H),2.32 (s, 3H), 2.2˜2.4(m, 2H), 3.20 (s, 3H), 3.0˜3.3 (m, 2H), 3.45 (dd,J=2.0 & 12.9 Hz, 1H), 3.64 (dd, J=4.7 & 10.6 Hz, 1H), 4.04 (d, J=12.9Hz, 1H), 4.40 (t, J=10.6 Hz, 1H), 4.70 (dd, J=4.7 & 10.6 Hz, 1H), 4.72(bs, 2H)

EXAMPLE 30 Synthesis of Compound 30

463 mg of Compound e obtained in REFERENCE EXAMPLE 5 was dissolved in 30ml of anhydrous tetrahydrofuran. While adding suspension of the sodiumsalt prepared from 152 mg of benzoylacetone and 48.4 mg of sodiumhydride (60% content, oil dispersion) in 10 ml of tetrahydrofuran inseveral portions, the solution was stirred at ambient temperature for 15minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=40:1 to 20:1) to give a violet fraction. After thesolvent was distilled off from this fraction, n-hexane-chloroform wasadded to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 47.9 mg (yield, 7.7%) of1aN-allyloxycarbonyl derivative of Compound 30 as a violet powder.

Subsequently product obtained above was dissolved in 2 ml of anhydroustetrahydrofuran, and 70 μl of triethylammonium formate and 8.1 mg oftetrakis (triphenylphosphine) palladium (0) were added to the solution.The mixture was stirred at ambient temperature for 2 hours and 10minutes under an argon atmosphere.

The reaction mixture was purified directly by column chromatography(silica gel; chloroform:methanol=20:1) to give a violet fraction. Thisfraction was then treated as described above to give 11.7 mg (yield,69%) of Compound 30 as a violet powder. TLC: Rf=0.37(chloroform:methanol=9:1)

FAB-MS (m/z): 479 (M⁺ +2); C₂₅ H₂₃ N₃ O₇ =477

IR (cm⁻¹): 3400, 3330, 3240, 2900, 1750, 1690, 1660, 1600, 1480, 1390,1360, 1240, 1230, 1220, 1190, 1100

¹ H-NMR (270 MHz, chloroform -d₁) δ (ppm): 0.66 (bs, 1H), 1.87 (s, 3H),2.85 (bs, 1H), 2.93 (bd, J=4.0 Hz, 1H), 3.15 (d, J=20.3 Hz, 1H), 3.24(s, 3H), 3.27 (d, J=20.3 Hz, 1H), 3.49 (dd, J=1.8 & 12.8 Hz, 1H), 3.69(dd, J=4.7 & 10.2 Hz, 1H), 4.07 (d, J=12.8 Hz, 1H), 4.61 (t, J=10.6 Hz,1H), 4.7˜4.8 (br, 2H), 4.73 (dd, J=4.7 & 10.7 Hz, 1H), 7.44˜7.51 (m,2H), 7.55˜7.62(m, 1H), 7.78˜7.83 (m, 2H)

EXAMPLE 31 Synthesis of Compound 31

400 mg of Compound f obtained in REFERENCE EXAMPLE 6 was dissolved in 30ml of anhydrous tetrahydrofuran. While adding a suspension of the sodiumsalt prepared from 150 μl of benzoylacetone and 48.9 mg of sodiumhydride (60% content, oil dispersion) in 10 ml of tetrahydrofuran inseveral portions, the solution was stirred at ambient temperature for anhour.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give a violet fraction. After the solventwas distilled off from this fraction, n-hexane-chloroform was added togive a powder. The solvent was distilled off and the residue wasthoroughly dried in vacuo to give 88.7 mg (yield, 18%) of Compound 31 asa violet powder.

TLC: Rf=0.57 (chloroform:methanol=9:1)

FAB-MS (m/z): 492 (M⁺ +1), 494 (M⁺ +3); C₂₆ H₂₅ N₃ O₇ =491

IR (cm⁻¹): 3400, 3300, 2880, 1760, 1740, 1690, 1660, 1610, 1480, 1390,1360, 1240, 1190, 1110, 1040

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 1.87 (s, 3H), 2.28 (s, 3H),2.2˜2.4 (m, 2H), 3.21 (s, 3H), 3.1˜3.3 (2H, 9a-overlapped with 9amethoxy), 3.44 (dd, J=1.7 & 12.8 Hz, 1H), 3.66 (dd, J=4.7 & 10.7 Hz,1H), 4.04 (d, J=12.8 Hz, 1H), 4.42 (t, J=10.7 Hz, 1H), 4.72 (dd, J=4.7 &10.7 Hz, 1H), 4.78 (bs, 2H), 7.44˜7.53 (m, 2H), 7.55˜7.62 (m, 1H),7.78˜7.83(m, 2H)

EXAMPLE 32 Synthesis of Compound 32

405 mg of Compound f obtained in REFERENCE EXAMPLE 6 was dissolved in 30ml of anhydrous tetrahydrofuran. While adding a suspension of the sodiumsalt prepared from 130 μl of ethyl acetoacetate and 51.5 mg of sodiumhydride (60% content, oil dispersion) in 10 ml of tetrahydrofuran inseveral portions, the solution was stirred at ambient temperature for 50minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=100:1) to give a violet fraction. After the solventwas distilled off from this fraction, n-hexane-chloroform was added togive a powder. The solvent was distilled off and the residue wasthoroughly dried in vacuo to give 59.9 mg (yield, 13%) of Compound 32 asa violet powder.

TLC: Rf=0.34 (chloroform:acetonitrile:n-hexane=5:5:1)

FAB-MS (m/z): 460 (M⁺ +1); C₂₂ H₂₅ N₃ O₈ =459

IR (cm⁻¹): 3450, 3350, 2950, 1720, 1710, 1700, 1640, 1630, 1580, 1570,1450, 1360, 1330, 1310, 1210, 1190, 1090

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 1.32 (t, J=6.9 Hz, 3H), 2.27(s, 3H), 2.20˜2.35 (m, 2H), 2.39(s, 3H), 3.08 (d, J=20.4 Hz, 1H), 3.20(d, J=20.4 Hz, 1H), 3.20 (s, 3H), 3.45 (dd, J=2.0 & 12.9 Hz, 1H), 3.63(dd, J=4.6 & 10.9 Hz, 1H), 4.04 (d, J=12.9 Hz, 1H), 4.23 (q, 2H), 4.41(bt, J=10.7 Hz, 1H), 4.70 (dd, J=4.6 & 10.6 Hz, 1H), 4.76 (bs, 2H)

EXAMPLE 33 Synthesis of Compound 34

1.00 g of Compound c obtained in REFERENCE EXAMPLE 3 was dissolved in 50ml of anhydrous tetrahydrofuran and 0.20 ml of anhydrous triethylamineand 372 mg of dimedone were added to the solution followed by stirringat ambient temperature for 2 hours and 15 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=30:1) to give a violet fraction.

After the solvent was distilled off from this fraction,n-hexane-chloroform was added to give a powder. The solvent wasdistilled off and the residue was thoroughly dried in vacuo to give276.5 mg (yield, 31% based on7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin B) ofCompound 34 as a violet powder.

TLC: Rf=0.32 (chloroform:methanol=9:1)

FAB-MS (m/z): 458 (M⁺ +3): C₂₃ H₂₅ N₃ O₇ =455

IR (cm⁻¹): 3470, 3420, 3300, 2950, 1710, 1660, 1630, 1590, 1390, 1350,1340, 1200, 1190, 1110

¹ H-NMR (270 MHz, pyridine-d₅) δ(ppm): 0.93 (s, 3H), 0.96 (s, 3H), 2.17(s, 3H), 2.2˜2.4 (m, 5H), 2.50 (d, J=4.6 Hz, 1H), 3.01 (d, J=20.5 Hz,1H), 3.10 (d, J=20.5 Hz, 1H), 3.57 (dd, J=1.7 & 12.7 Hz, 1H), 4.13 (d,J=12.7 Hz, 1H), 4.26 (dd, J=3.3 & 9.1 Hz, 1H), 5.25 (dd, J=9.1 & 10.7Hz, 1H), 5.43 (dd, J=3.3 & 10.7 Hz, 1H), 7.3˜7.7 (br, 2H), 8.34 (br, 1H)

EXAMPLE 34 Synthesis of Compound 35

732 mg of Compound c obtained in REFERENCE EXAMPLE 3 was dissolved in 50ml of anhydrous tetrahydrofuran and 0.20 ml of anhydrous triethylamineand 218 mg of 1,3-cyclohexanedione were added to the solution followedby stirring at ambient temperature for 45 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was purified by column chromatography (silica gel;chloroform:methanol=50:1 to 30:1) to give a violet fraction. After thesolvent was distilled off from this fraction, n-hexane-chloroform wasadded to give a powder. The solvent was distilled off and the residuewas thoroughly dried in vacuo to give 151.4 mg (yield, 25% based on7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin B) ofCompound 35 as a violet powder.

TLC: Rf=0.35 (chloroform:methanol=9:1)

FAB-MS (m/z): 429 (M⁺ +2), 430 (M⁺ +3); C₂₁ H₂₁ N₃ O₇ =427

IR (cm⁻¹): 3450, 3200, 2950, 1710, 1660, 1620, 1570, 1450, 1380, 1350,1210, 1190, 1120, 1070, 1060

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 2.05 (m, 2H), 2.26 (s, 3H),2.28 (s, 2H), 2.45 (m, 2H), 2.61 (m, 2H), 2.95(d, J=20.8 Hz, 1H), 3.08(d, J=20.8 Hz, 1H), 3.45 (d, J=12.9 Hz, 1H), 3.76 (t, J=4.1 Hz, 1H),4.00 (d, J=12.9 Hz, 1H), 4.62 (bs, 1H), 4.72 (d, J=4.1 Hz, 2H), 4.76(bs, 2H)

EXAMPLE 35 Synthesis of Compound 36 and Compound 37

After 568 mg of Compound e obtained in REFERENCE EXAMPLE 5 was dissolvedin 40 ml of anhydrous tetrahydrofuran, 0.20 ml of anhydroustriethylamine and 157 mg of 5-methyl-1,3-cyclohexanedione were added tothe solution. The mixture was stirred at ambient temperature for an hourand 15 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was again dissolved in 100 ml of chloroform. 100 ml ofsilica gel was added to the solution to adsorb the product and allowedto stand at ambient temperature for 20 hours and 30 minutes.

The silica gel was eluted with an eluent (chloroform:methanol=9:1). Theresulting violet solution was concentrated under reduced pressure andthe residue was purified by column chromatography (silica gel;chloroform:methanol=50:1) to give a violet fraction. After the solventwas distilled off from this fraction, n-hexane-chloroform was added togive a powder. The solvent was distilled off and the residue wasthoroughly dried in vacuo to give 405.1 mg (yield, 62%) of the1aN-allyloxycarbonyl derivatives of Compounds 36 and 37 as a violetpowder.

Subsequently, 405.0 mg of the product obtained above was dissolved in 20ml of anhydrous tetrahydrofuran, and 0.40 ml of triethylammonium formateand 42 mg of tetrakis (triphenylphosphine) palladium (0) was added tothe solution. The mixture was stirred at ambient temperature for 35minutes under an argon atmosphere.

The reaction mixture was purified directly by column chromatography(silica gel; chloroform:methanol=50:1 to 20:1) to give a violetfraction. This fraction was then treated as described above to give222.9 mg (yield, 66%) of the mixture of Compounds 36 and 37 as a violetpowder. Each diastereoisomer was isolated and purified by preparativeHPLC to give 100.5 mg of a diastereoisomer (Compound 36) having ashorter retention time and 74.3 mg of another diastereoisomer (Compound37) having a longer retention time.

Compound 36

TLC: Rf=0.37 (chloroform:methanol 9:1)

HPLC: t_(R) =20.83 minutes (eluent:acetonitrile:water=25:75; column: YMCAM-312 S-5; flow rate: 1.0 ml/min)

FAB-MS (m/z): 442(M⁺ +1); C₂₂ H₂₃ N₃ O₇ =441

IR(cm⁻¹): 3450, 3350, 3300, 3200, 2960, 2890, 1720, 1660, 1630, 1570,1450, 1390, 1340, 1200, 1190, 1160, 1130, 1070, 1030

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.5˜1.0 (br, 1H), 1.11 (d,J=5.9 Hz, 3H), 2.13 (dd, J=12.1 & 16.0 Hz, 1H), 2.2˜2.4 (m, 2H), 2.54(dd, J=2.5 & 16.0 Hz, 1H), 2.66 (m, 1H), 2.85 (bd, J=4.5 Hz, 1H),2.92(d, J=4.5 Hz, 1H), 2.94(d, J=20.8 Hz, 1H), 3.12 (d, J=20.8 Hz, 1H),3.22(s, 3H), 3.49 (dd, J=1.5 & 12.9 Hz, 1H), 3.67 (dd, J=4.6 & 10.4 Hz,1H), 4.07 (d, J=12.9 Hz, 1H), 4.58 (bt, J=10.6 Hz, 1H), 4.72 (dd, J=4.7& 10.6 Hz, 1H), 4.77 (bs, 2H)

Compound 37

TLC: Rf=0.37 (chloroform:methanol=9:1)

HPLC: t_(R) =22.60 minutes (eluent:acetonitrile:water=25.75; column: YMCAM-312 S-5; flow rate: 1.0 ml/min)

FAB-MS (m/z): 442 (M⁺ +1); C₂₂ H₂₃ N₃ O₇ =441

IR (cm⁻¹): 3450, 3350, 3300, 3200, 2960, 2890, 1720, 1660, 1630, 1570,1450, 1390, 1340, 1200, 1190, 1160, 1130, 1070, 1030

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.5˜1.0 (br, 1H), 1.12(d, J=5.9Hz, 3H), 2.14 (dd, J=12.4 & 15.8 Hz, 1H), 2.2˜2.4 (m, 2H), 2.54 (dd,J=2.0 & 15.8 Mz, 1H), 2.66(m, 1H), 2.85 (dd, J=2.0 & 4.5 Hz, 1H), 2.92(d, J=4.5 Hz, 1H), 2.99 (d, J=20.8 Hz, 1H), 3.08(d, J=20.8 Hz, 1H), 3.22(s, 3H), 3.49 (dd, J=1.5 & 12.9 Hz, 1H), 3.67 (dd, J=4.7 & 10.1 Hz, 1H),4.07 (d, J=12.9 Hz, 1H), 4.58 (bt, J=10.4 Hz, 1H), 4.71 (dd, J=4.7 &10.6 Hz, 1H), 4.7˜4.8 (bs, 2H)

EXAMPLE 36 Synthesis of Compound 38 and Compound 39

After 928 mg of Compound e obtained in REFERENCE EXAMPLE 5 was dissolvedin 70 ml of dichloromethane, 0.20 ml of anhydrous triethylamine and 437mg of 5-(p-methoxyphenyl)-1,3-cyclohexanedione were added to thesolution. The mixture was stirred at ambient temperature for 50 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was again dissolved in 200 ml of chloroform. 100 ml ofsilica gel was added to the solution to adsorb the product and allowedto stand at ambient temperature for 17 hours.

The silica gel was eluted with an eluent (chloroform:methanol=9:1). Theresulting violet solution was concentrated under reduced pressure andthe residue was purified by column chromatography (silica gel;chloroform:methanol=100:1) to give a violet fraction. After the solventwas distilled off from this fraction, n-hexane-chloroform was added togive a powder. The solvent was distilled off and the residue wasthoroughly dried in vacuo to give 923.4 mg (yield, 73%) of the1aN-allyloxycarbonyl derivatives of Compounds 38 and 39 as a violetpowder.

Subsequently, 802.5 mg of the product obtained above was dissolved in 40ml of anhydrous tetrahydrofuran, and 1.0 ml of triethylammonium formateand 43 mg of tetrakis (triphenylphosphine) palladium (0) were added tothe solution. The mixture was stirred at ambient temperature for 25minutes under an argon atmosphere.

The reaction mixture was purified directly by column chromatography(silica gel; chloroform:methanol=50:1 to 20:1) to give a violetfraction. This fraction was then treated as described above to give126.6 mg (yield, 19%) of the mixture of Compounds 38 and 39 as a violetpowder. Each diastereoisomer was isolated and purified by preparativeHPLC to give 44.5 mg of a diastereoisomer (Compound 38) having a shorterretention time and 34.2 mg of another diastereoisomer (Compound 39)having a longer retention time.

Compound 38

TLC: Rf=0.38 (chloroform:methanol=9:1)

HPLC: t_(R) =9.66 minutes (eluent:acetonitrile:water=40:60; column: YMCAM-312 S-5; flow rate: 1.0 ml/min)

FAB-MS (m/z): 534 (M⁺ +1); C₂₈ H₂₇ N₃ O₈ =533

IR (cm⁻¹): 3500, 3400, 3300, 3180, 2920, 1720, 1660, 1630, 1580, 1510,1390, 1340, 1250, 1200, 1160, 1120, 1070, 1030

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.8˜1.0 (br, 1H), 2.61 (dd,J=12.4 & 16.3 Hz, 1H), 2.7˜3.0 (m, 4H), 2.93 (d, J=4.5 Hz, 1H), 2.99 (d,J=20.3 Hz, 1H), 3.17 (d, J=20.3 Hz, 1H), 3.22 (s, 3H), 3.3˜3.4(m, 1H),3.50 (dd, J=2.0 & 12.9 Hz, 1H), 3.67 (dd, J=4.6 & 10.1 Hz, 1H), 3.80 (s,3H), 4.08 (d, J=12.9 Hz, 1H), 4.59 (bt, J=10.4 Hz, 1H), 4.72 (dd, J=4.6& 10.9 Hz, 1H), 4.7˜4.8 (br, 2H), 6.88 (d, J=8.4 Hz, 2H), 7.15 (d, J=8.4Hz, 2H)

Compound 39

TLC: Rf=0.38 (chloroform:methanol=9:1)

HPLC: t_(R) =10.40 minutes (eluent:acetonitrile:water=40:60; column: YMCAM-312 S-5; flow rate: 1.0 ml/min)

FAB-MS (m/z): 534 (M⁺ +1); C₂₈ H₂₇ N₃ O₈ =533

IR (cm⁻¹): 3500, 3400, 3300, 3180, 2920, 1720, 1660, 1650, 1630, 1580,1570, 1520, 1390, 1340, 1250, 1200, 1070, 1030

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.8˜1.0 (br, 1H), 2.62 (dd,J=12.4 & 16.1 Hz, 1H), 2.77 (dd, J=4.5 & 16.1 Hz, 1H), 2.7˜2.9 (m, 3H),2.92 (d, J=5.0 Hz, 1H), 3.04 (d, J=20.3 Hz, 1H), 3.13 (d, 20.3 Hz, 1H),3.23 (s, 3H), 3.3˜3.4 (m, 1H), 3.50 (dd, J=2.0 & 12.9 Hz, 1H), 3.68 (dd,J=4.8 & 10.4 Hz, 1H), 3.81 (s, 3H), 4.08 (d, J=12.9 Hz, 1H), 4.59 (bt,J=10.4 Hz, 1H), 4.71 (dd, J=4.8 & 10.6 Hz, 1H), 4.7˜4.8 (br, 2H), 6.88(d, J=8.4 Hz, 2H), 7.15(d, J=8.4 Hz, 2H)

EXAMPLE 37 Synthesis of Compound 40

After 932 mg of Compound e obtained in REFERENCE EXAMPLE 5 was dissolvedin 70 ml of dichloromethane, 0.20 ml of anhydrous triethylamine and 520mg of 5-(2,6-dichlorophenyl)-1,3-cyclohexanedione were added to thesolution. The mixture was stirred at ambient temperature for 40 minutes.

The reaction mixture was subjected to work-up as in EXAMPLE 28. Theresidue obtained was again dissolved in 200 ml of chloroform. 100 ml ofsilica gel was added to the solution to adsorb the product and allowedto stand at ambient temperature for 21 hours.

The silica gel was eluted with an eluent (chloroform:methanol=9:1). Theresulting violet solution was concentrated under reduced pressure andthe residue was purified by column chromatography (silica gel;chloroform:methanol=100:1) to give a violet fraction. After the solventwas distilled off from this fraction, n-hexane-chloroform was added togive a powder. The solvent was distilled off and the residue wasthoroughly dried in vacuo to give 601.0 mg (yield, 47%) of the1aN-allyloxycarbonyl derivative of Compound 40 as a violet powder.

Subsequently, 591.5 mg of the product obtained above was dissolved in 30ml of anhydrous tetrahydrofuran, and 0.50 ml of triethylammonium formateand 31 mg of tetrakis (triphenylphosphine) palladium (0) were added tothe solution. The mixture was stirred at ambient temperature for 23minutes under an argon atmosphere.

The reaction mixture was purified directly by column chromatography(silica gel; chloroform:methanol=20:1) to give a violet fraction. Thisfraction was then treated as described above to give 35.3 mg (yield, 7%)of Compound 40 as a violet powder.

TLC: Rf=0.49 (chloroform:methanol=9:1)

FAB-MS(m/z): 571, 573, 575 (M⁺ +1); C₂₇ H₂₃ ³⁵ C1₂ N₃ O₇ =570

IR (cm⁻¹): 2500, 3400, 3300, 3200, 2920, 1720, 1640, 1630, 1580, 1570,1440, 1390, 1340, 1200, 1080

¹ H-NMR (270 MHz, chloroform-d₁) δ(ppm): 0.8˜0.9 (br, 1H), 2.52 (dd,J=4.2 & 17.1 Hz, 1H), 2.66 (dd, J=4.2 & 17.1 Hz, 1H), 2.86 (dd, J=2.0 &4.5 Hz, 1H), 2.93 (d, J=4.5 Hz, 1H), 3.08 (bd, J=16.8 Hz, 1H), 3.14 (bd,J=16.8 Hz, 1H), 3.23(s, 3H), 3.51 (dd, J=2.0 & 12.9 Hz, 1H),3.45˜3.71(m, 2H), 3.68 (dd, J=4.5 & 10.4 Hz, 1H), 4.09 (d, J=12.9 Hz,1H), 4.3˜4.5(m, 1H), 4.59 (t, J=10.6 Hz, 1H), 4.71 (dd, J=4.5 & 10.9 Hz,1H), 4.7˜4.8 (br, 2H), 7.16 (t, J=8.2 Hz, 1H), 7.33 (m, 2H)

Hereafter the structures of the Compounds in REFERENCE EXAMPLES aregiven.

                                      TABLE 6    __________________________________________________________________________     ##STR17##    Compound             A   R.sup.1 R.sup.2 Y  Z    __________________________________________________________________________    a       CH.sub.2                 CH.sub.2 OCONH.sub.2                         H       CH.sub.3                                    COCH.sub.3    b       SePh CH.sub.2 OCONH.sub.2                         H       CH.sub.3                                    COCH.sub.3    c       CH.sub.2                 H       CH.sub.2 OCONH.sub.2                                 H  CH.sub.3             SePh    d            CH.sub.2 OCONH.sub.2                         H       CH.sub.3                                    CO.sub.2 CH.sub.2 CHCH.sub.2            CH.sub.3    e       CH.sub.2                 CH.sub.2 OCONH.sub.2                         H       CH.sub.3                                    CO.sub.2 CH.sub.2 CHCH.sub.2    f       CH.sub.2                 CH.sub.2 OCONH.sub.2                         H       CH.sub.3                                    CH.sub.3    __________________________________________________________________________     *SePh represents phenylseleno.

REFERENCE EXAMPLE 1 Synthesis ofIa-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-methylenemitomycinA (Compound a)

After 5.77 g ofIa-acetyl-7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycinA (described in JP-A 70490/89) was dissolved in 100 ml ofdichloromethane, 2.82 g of potassium carbonate was added to thesolution. Then a solution of 2.59 g of m-chloroperbenzoic acid (purityabout 80%) in 50 ml of dichloromethane was added dropwise to thesolution at -40° C. over 15 minutes. Subsequently the mixture wasstirred at -40° to -30° C. for 40 minutes and then at 20° C. for further50 minutes. The reaction mixture was washed with a mixture (1:1) ofaqueous sodium thiosulfate solution (M/10) and saturated aqueous sodiumhydrogencarbonate solution and further with saturated aqueous sodiumchloride solution. After drying over anhydrous sodium sulfate, themixture was concentrated under reduced pressure. n-Hexane was added tothe residue to give a powder. The obtained powder was filtered andthoroughly dried in vacuo to give 3.77 g (yield, 90%) of Compound a. Thepurity of Compound a was about 90% by ¹ H-NMR.

TLC: Rf=0.42 (chloroform:acetonitrile:n-hexane=5:5:1)

FAB-MS (m/z): 420 (M⁺ +1); C₁₉ H₂₁ N₃ O₈ =419

¹ H-NMR (400 MHz, chloroform-d₁ /pyridine-d₅) δ(ppm): 2.08 (s, 3H), 3.17(s, 3H), 3.24 (dd, J=4.4 & 2.0 Hz, 1H), 3.50(d, J=4.4 Hz, 1H), 3.48 (dd,J=13.3 & 2.0 Hz, 1H), 3.79 (dd, J=11.1 & 4.9 Hz, 1H), 4.17 (t, J=10.8Hz, 1H), 4.04˜4.29(m, 4H), 4.38(d, J=13.3 Hz, 1H), 5.04(dd, J=10.8 & 4.7Hz, 1H), 5.79 (bs, 2H), 6.09 (bs, 1H), 6.36 (bs, 1H)

REFERENCE EXAMPLE 2 Synthesis ofIa-acetyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycinA (Compound b)

After 421 mg of Compound a obtained in REFERENCE EXAMPLE 1 was dissolvedin 25 ml of anhydrous tetrahydrofuran, 238 mg of N-(phenylselenenyl)morphotine was added to the solution. The mixture was stirred at 25° C.for 3 hours. The reaction mixture was purified directly by columnchromatography (silica gel; chloroform:methanol=30:1) to give a yellowfraction. The solvent was distilled off from this fraction and theresidue was dissolved in a small amount of chloroform. n-Hexane wasadded to give a powder. The powder was filtered and thoroughly dried invacuo to give 318 mg (yield, 56%) of Compound b as a yellow powder.Compound b was the equilibrium mixture of diastereoisomer based on theconfiguration at the 6-position.

TLC: Rf=0.30 (chloroform-acetonitrile-n-hexane=5:5:1)

FAB-MS (m/z): 562, 564 (M⁺ +1); C₂₄ H₂₅ N₃ O₈ ⁷⁸ Se=561

¹ H-NMR (400 MHz, chloroform-d₁) δ(ppm):

major diastereoisomer: 2.20 (s, 3H), 3.21 (s, 3H), 3.26 (dd, J=2.0 & 4.7Hz, 1H), 3.41 (dd, J=2.0 & 13.0 Hz, 1H), 3.52 (d, J=4.7 Hz, 1H), 3.73(dd, =4.9 & 10.8 Hz, 1H), 3.84 (d, J=13.0 Hz, 1H), 4.02 (s, 1H), 4.19(t, J=11 Hz, 1H), 4.01˜4.20 (m, 3H), 4.41 (m, 1H), 4.91 (bs, 2H), 4.95(dd, J=4.9 & 11.1 Hz, 1H), 7.28˜7.38 (m, 3H), 7.61(m, 2H)

minor diastereoisomer: 2.10 (s, 3H), 3.21 (s, 3H), 3.23 (dd, J=2.0 & 4.4Hz, 1H), 3.39 (dd, J=2.0 & 13.0 Hz, 1H), 3.48 (d, J=4.4 Hz, 1H), 3.67(dd, J=4.7 & 10.8 Hz, 1H), 4.17 (s, 1H), 4.01˜4.20 (m, 4H), 4.31 (m,1H), 4.40 (d, J=13.0 Hz, 1H), 4.81 (dd, J=4.7 & 10.8 Hz, 1H), 4.89 (bs,2H), 7.28˜7.38 (m, 3H), 7.61 (m, 2H)

REFERENCE EXAMPLE 3 Synthesis of7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-methylenemitomycinB (Compound c)

After 1.25 g of7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin A(described in JP-A 70490/89) was dissolved in 25 ml of dichloromethane,650 mg of potassium carbonate was added to the solution. Then a solutionof 630 mg of m-chloroperbenzoic acid (purity about 80%) in 15 ml ofdichloromethane was added dropwise to the solution at -40° C. over 15minutes. Subsequently the mixture was stirred at -30° C. for 35 minutesand then at 20° C. for further 40 minutes. The reaction mixture wasfiltered through filter aide (dialomucous earth). After the filtrate wasconcentrated under reduced pressure, n-hexane was added to the residueto give a powder. The powder was filtered and thoroughly dried in vacuoto give 1.05 g of the crude product of Compound c as a yellow powder.

TLC: Rf=0.30 (chloroform:methanol=9:1)

REFERENCE EXAMPLE 4 Synthesis ofIa-allyloxycarbonyl-7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycinA (Compound d)

After 1.31 g ofIa-allyloxycarbonyl-7-demethoxy-7-ethylenedioxy-6,7-dihydromitomycin A(described in JP-A 49786/90) was dissolved in 30 ml of anhydrousacetonitrile and 1.0 ml of triethylamine, a solution of 1.01 g ofphenylselenenyl bromide in 10 ml of anhydrous acetonrile was addeddropwise to the solution at 0° C. over 5 minutes. The mixture wasstirred at ambient temperature for 14 hours. The reaction mixture wasdiluted with a phosphate buffer (M/20, pH 7) followed by extraction withchloroform. The organic layer obtained was washed with saturated aqueousammonium chloride solution and saturated aqueous sodium chloridesolution successively. After drying over anhydrous sodium sulfate, thesolvent was distilled off under reduced pressure. The residue waspurified by column chromatography (silica gel;chloroform:methanol=100:1) to give a yellow fraction. The solvent wasdistilled off from this fraction and the residue was dissolved in asmall amount of chloroform. n-Hexane was added to give a powder. Thepowder was filtered and thoroughly dried in vacuo to give 1.36 g (yield,78%) of the desired compound as a yellow powder. The product was themixture of diastereoisomers based on the configuration at the6-position.

TLC: Rf=0.54 (chloroform:methanol=15:1)

¹ H-NMR (90 MHz, chloroform-d₁) δ(ppm):

major diastereoisomer: (major peaks) 1.54 (s, 3H), 3.18 (s, 3H), 5.24(dd, J=2 & 9 Hz, 1H), 5.31 (dd, J=2 & 18 Hz, 1H), 5.6-6.2 (m, 1H),7.2-7.8 (m, 5H)

minor diastereoisomer: (major peaks) 1.40 (s, 3H), 3.28 (s, 3H)

REFERENCE EXAMPLE 5 Synthesis ofIa-allyloxycarbonyl-7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-methylenemitomycinA (Compound e)

After Compound d (3.89 g) obtained in REFERENCE EXAMPLE 4 was dissolvedin 70 ml of dichloromethane, 2.62 g of potassium carbonate was added tothe solution. Then a solution of 2.13 g of m-chloroperbenzoic acid(purity about 80%) in 40 ml of dichloromethane was added dropwise to thesolution at -40° C. over 10 minutes. Subsequently the mixture wasstirred at -40° C. for 30 minutes and then at 20° C. for further anhour. The reaction mixture was washed with a mixture (1:1) of aqueoussodium thiosulfate solution (M/10) and saturated aqueous sodiumhydrogencarbonate solution and further with saturated aqueous sodiumchloride solution. After drying over anhydrous sodium sulfate, themixture was concentrated under reduced pressure. n-Hexane was added tothe residue to give a powder. The obtained powder was filtered andthoroughly dried in vacuo to give 2.25 g (yield, 78%) of Compound e. Thepurity of Compound e was about 90% by ¹ H-NMR.

TLC: Rf=0.66 (chloroform:methanol=9:1)

¹ H-NMR (90 MHz, chloroform-d₁) δ(ppm): (major peaks) 3.22 (s, 3H), 5.26(dd, J=2 & 11 Hz, 1H), 5.30 (dd, J=2 & 18 Hz, 1H), 6.11 (s, 1H), 6.38(s, 1H)

REFERENCE EXAMPLE 6 Synthesis of7-demethoxy-6-demethyl-7-ethylenedioxy-6,7-dihydro-6-methylenemitomycinF (Compound f)

After 2.74 g of7-demethoxy-7-ethylenedioxy-6,7-dihydro-6-phenylselenomitomycin F(described in JP-A 167282/90) was dissolved in 100 ml ofdichloromethane, 2.09 g of potassium carbonate was added to thesolution. While stirring at -40° C., a solution of 1.39 g ofm-chloroperbenzoic acid (purity about 80%) in 50 ml of dichloromethanewas added dropwise to the solution over 15 minutes. Subsequently themixture was stirred at -40° to -20° C. for 40 minutes and then at 20° C.for further an hour and 50 minutes.

The reaction mixture was washed with a mixture (1:1) of aqueous sodiumthiosulfate solution (M/10) and saturated aqueous sodiumhydrogencarbonate solution and further with saturated aqueous sodiumchloride solution. After drying over anhydrous sodium sulfate, themixture was concentrated under reduced pressure. n-Hexane was added tothe residue to give a powder. The powder obtained was filtered andthoroughly dried in vacuo to give 1.73 g (yield, 89%) of Compound f. Thepurity of Compound f was about 90% by ¹ H-NMR.

TLC: Rf=0.51 (chloroform:methanol=9:1)

FAB-MS: 392 (M⁺ +1); C₁₈ H₂₁ N₃ O₇ =391

IR (cm⁻¹): 3450, 3370, 2960, 1720, 1660, 1570, 1450, 1400, 1360, 1340,1210, 1140, 1120, 1060

¹ H-NMR (400 MHz, chloroform-d₁) δ(ppm): 2.24 (dd, J=2.2 & 4.7 Hz, 1H),2.26 (s, 3H), 2.30 (d, J=4.7 Hz, 1H), 3.19 (s, 3H), 3.44 (dd, J=2.2 &12.6 Hz, 1H), 3.61 (dd, J=4.7 & 10.6 Hz, 1H), 4.15 (d, J=12.6 Hz, 1H),4.06˜4.34 (m, 4H), 4.40 (t, J=10.6 Hz, 1H), 4.73 (bs, 2H), 4.74 (dd,J=4.7 & 10.6 Hz, 1H), 6.07 (d, J=1.5 Hz, 1H), 6.34 (d, J=1.5 Hz, 1H)

According to the present invention, novel mitomycin derivatives havingan excellent antitumor activity are provided.

What is claimed is:
 1. A mitomycin derivative represented by theformula: ##STR18## wherein U and V independently represent a substitutedor unsubstituted lower alkyl having 1 to 6 carbon atoms, lower alkenylhaving 2 to 6 carbon atoms or lower alkoxy group having 1 to 6 carbonatoms, a substituted or unsubstituted phenyl, naphthyl or loweralkenyloxy group having 2 to 6 carbon atoms, or U and V together form--(CH₂)_(m) --CR³ R⁴ --CH₂ --, wherein m represents an integer of 0 to1, R³ and R⁴ independently represent hydrogen, a lower alkyl having 1 to6 carbon atoms or a substituted or unsubstituted phenyl or naphthyl; Yrepresents hydrogen or methyl; Z represents hydrogen, methyl, a loweralkanoyl having 2 to 7 carbon atoms or an allyloxycarbonyl; one of R¹and R² represents CH₂ OCONH₂ and the other represents hydrogen, or R¹and R² together form methylene (═CH₂)wherein the substituent on therespective groups is lower alkoxy having 1 to 6 carbon atoms, loweralkanoyl having 2 to 7 carbon atoms, alkoxycarbonyl having 2 to 7 carbonatoms, aroyl having 7 to 11 carbon atoms, oxo, halogen, alkanoylaminohaving 2 to 7 carbon atoms, or nitro.
 2. The mitomycin derivative asclaimed in claim 1, wherein Y is methyl;Z is hydrogen; R¹ is CH₂ OCONH₂and R² is hydrogen.
 3. The mitomycin derivative as claimed in claim 2,wherein U ismethyl; and V is ethoxy.
 4. The mitomycin derivative asclaimed in claim 2, wherein U isphenyl; and V is methyl.
 5. Themitomycin derivative as claimed in claim 2, wherein m is
 1. 6. Themitomycin derivative as claimed in claim 5, wherein R³ is hydrogen. 7.The mitomycin derivative as claimed in claim 6, wherein R⁴ is hydrogen.8. The mitomycin derivative as claimed in claim 6, wherein R⁴ is a loweralkyl having 1 to 6 carbon atoms.
 9. The mitomycin derivative as claimedin claim 8, wherein R⁴ is methyl.
 10. The mitomycin derivative asclaimed in claim 6, wherein R⁴ is a substituted or unsubstituted phenyl.11. The mitomycin derivative as claimed in claim 10, wherein R⁴ isp-methoxyphenyl.
 12. The mitomycin derivative as claimed in claim 5,wherein R³ is a lower alkyl having 1 to 6 carbon atoms; and R⁴ is alower alkyl having 1 to 6 carbon atoms.
 13. The mitomycin derivative asclaimed in claim 12, wherein R³ ismethyl; and R⁴ is methyl. 14.Pharmaceutical composition comprising a compound according to any one ofclaims 1, 2 or 3-13, inclusive as the active ingredient and apharmaceutically acceptable carrier.